Modulators of Cystic Fibrosis Transmembrane Conductance Regulator

ABSTRACT

The present invention relates to modulators of cystic fibrosis Transmembrane Conductance Regulator (“CFTR”), compositions thereof, and methods therewith. The present invention also relates to methods of treating CFTR mediated diseases using such modulators.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present invention claims priority under 35 U.S.C. §119 to U.S.Provisional Application Ser. No. 60/707,380, filed Aug. 11, 2005, andU.S. Provisional Application Ser. No. 60/732,476, filed Nov. 2, 2005.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to modulators of cystic fibrosisTransmembrane Conductance Regulator (“CFTR”), compositions thereof, andmethods therewith. The present invention also relates to methods oftreating CFTR mediated diseases using such modulators.

BACKGROUND OF THE INVENTION

ABC transporters are a family of membrane transporter proteins thatregulate the transport of a wide variety of pharmacological agents,potentially toxic drugs, and xenobiotics, as well as anions. ABCtransporters are homologous membrane proteins that bind and use cellularadenosine triphosphate (ATP) for their specific activities. Some ofthese transporters were discovered as multidrug resistance proteins(like the MDR1-P glycoprotein, or the multidrug resistance protein,MRP1), defending malignant cancer cells against chemotherapeutic agents.To date, 48 ABC Transporters have been identified and grouped into 7families based on their sequence, identity and function.

ABC transporters regulate a variety of important physiological roleswithin the body and provide defense against harmful environmentalcompounds. Because of this, they represent important potential drugtargets for the treatment of diseases associated with defects in thetransporter, prevention of drug transport out of the target cell, andintervention in other diseases in which modulation of ABC transporteractivity may fee beneficial.

One member of the ABC transporter family commonly associated withdisease is the cAMP/ATP-mediated anion channel, CFTR. CFTR is expressedin a variety of cells types, including absorptive and secretoryepithelia cells, where it regulates anion flux across the membrane, aswell as the activity of other ion channels and proteins. In epitheliacells, normal functioning of CFTR is critical for the maintenance ofelectrolyte transport throughout the body, including respiratory anddigestive tissue. CFTR is composed of approximately 1480 amino acidsthat encode a protein made up of a tandem repeate of transmembranedomains, each containing six transmembrane helices and a nucleotidebinding domain. The two transmembrane domains are linked by a large,polar, regulatory (R)-domain with multiple phosphorylation sites thatregulate channel activity and cellular trafficking.

The gene encoding CFTR has been identified and sequenced (See Gregory,R. J. et al, (1990) Nature 347:382-386; Rich, D. F. et at. (1990) Nature347:358-362), (Riordan, J, R. et al. (1989) Science 245:1066-1073). Adefect in this gene causes mutations in CFTR resulting in cysticfibrosis, the most common fatal genetic disease in humans. Cysticfibrosis affects approximately one in every 2,500 infants in the UnitedStates. Within the general United States population, up to 10 millionpeople carry a single copy of the detective gene without apparent illeffects. In contrast, individuals with two copies of the cystic fibrosisassociated gene suffer from the debilitating and fatal effects of cysticfibrosis, including chronic lung disease.

In patients with cystic fibrosis, mutations in CFTR endogenouslyexpressed in respiratory epithelia leads to reduced apical anionsecretion causing an imbalance in ion and fluid transport. The resultingdecrease in anion transport contributes to enhanced mucus accumulationin the lung and the accompanying microbial infections that ultimatelycause death in cystic fibrosis patients. In addition to respiratorydisease, cystic fibrosis patients typically suffer from gastrointestinalproblems and pancreatic insufficiency that, if left untreated, resultsin death. In addition, the majority of mates with cystic fibrosis areinfertile and fertility is decreased among females with cystic fibrosis.In contrast to the severe effects of two copies of the cystic fibrosisassociated gene, Individuals with a single copy of the cystic fibrosisassociated gene exhibit increased resistance to cholera and todehydration resulting from diarrhea—perhaps explaining the relativelyhigh frequency of the cystic fibrosis gene within the population.

Sequence analysis of the CFTR gene of cystic flbrosischromosomes hasrevealed a variety of disease causing mutations (Cutting, G. R. et al.(1990) Nature 346:366-369; Dean, M. et al. (1990) Cell 61:863:870; andKerem, B-S. et al. (1989) Science 245; 1073-1080; Kerem, B-S et al.(1990) Proc. Natl. Acad. Sci. USA 87:8447-8451). To date, >1000 diseasecausing mutations in the cystic fibrosis gene have been identified.(http://www.genet.sickkids.on.ca/cftr/). The most prevalent mutation isa deletion of phenylalanine at position 508 of the CFTR amino acidsequence, and is commonly referred to as ΔF508-CFTR. This mutationoccurs in approximately 70% of the cases of cystic fibrosis and isassociated with a severe disease.

The deletion of residue 508 in ΔP508-CFTR prevents the nascent proteinfrom folding correctly. This results in the inability of the mutantprotein to exit the endoplasmic reticulum (“ER”), and traffic to theplasma membrane. As a result, the number of channels present in themembrane is far less than observed in cells expressing wild-type CFTR.In addition to impaired trafficking, the mutation results in defectivechannel gating. Together, the reduced number of channels in the membraneand the defective gating lead to reduced anion transport acrossepithelia leading to defective ion and fluid transport. (Quinton, P. M.(1990), FASEB J. 4: 2709-2727). Studies have shown, however, that thereduced numbers of ΔF508-CFTR in the membrane are functional, albeitless than wild-type CFTR. (Dalemans et al. (1991), Nature Lond. 354:526-528; Denning et al., supra; Pasyk and Foskett (1995), J. Cell.Biochem. 270: 12347-50). In addition to ΔF508-CFTR, other diseasecausing mutations in CFTR that result in defective trafficking,synthesis, and/or channel gating could be up- or down-regulated to alteranion secretion and modify disease progression and/or severity.

Although CFTR transports a variety of molecules in addition to anions,it is clear that this role (the transport of anions) represents oneelement in an important mechanism of transporting ions and water acrossthe epithelium. The other elements include the epithelial Na⁺ channelENaC, Na⁺/2Cl⁻/K⁺ co-transporter, Na⁺-K⁺-ATPase pump and the basolateralmembrane K⁺ channels, that are responsible for the uptake of chlorideinto the cell.

These elements work together to achieve directional transport across theepithelium via their selective expression and localization within thecell. Chloride absorption lakes place by the coordinated activity ofENaC and CFTR present on the apical membrane and the Na⁺-K⁺-ATPasepump-and Cl− channels expressed on the basolateral surface of the cell.Secondary active transport of chloride from the luminal side leads tothe accumulation of intracellular chloride, which can then passivelyleave the cell via Cl⁻ channels, resulting in a vectorial transport.Arrangement of Na⁺/2Cl⁻/K⁺ co-transporter, Na⁺-K⁺-ATP ase pump and thebaselateral membrane K⁺ channels on the basolateral surface and CFTR onthe luminal side coordinate the secretion of chloride via CFTR on theluminal side. Because water is probably never actively transporteditself, its flow across epithelia depends on tiny transepithelialosmotic gradients generated by the bulk flow of sodium and chloride.

In addition to cystic fibrosis, modulation of CFTR activity may bebeneficial for other diseases not directly caused by mutations in CFTR,such as secretory diseases and other protein folding diseases mediatedby CFTR. These include, but are not limited to, chronic obstructivepulmonary disease (COFD), dry eye disease, and Sjögren's Syndrome.

COPD is characterized by airflow limitation that is progressive and notfully reversible. The airflow limitation is due to mucus hypersecretion,emphysema, and bronchiolitis. Activators of mutant or wild-type CFTRoffer a potential treatment of mucus hypersecretion and impairedmucociliary clearance that is common in COPD. Specifically, increasinganion secretion across CFTR may facilitate fluid transport into theairway surface liquid to hydrate the mucus and optimized periciliaryfluid viscosity. This would lead to enhanced mucociliary clearance and areduction in the symptoms associated with COPD. Dry eye disease ischaracterised by a decrease in tear aqueous production and abnormal tearfilm lipid, protein and mucin profiles. There are many causes of dryeye, some of which include age, Lasik eye surgery, arthritis,medications, chemical/thermal burns, allergies, and diseases, such ascystic fibrosis and Sjögrens's syndrome. Increasing anion secretion viaCFTR would enhance fluid transport from the corneal endothelial cellsand secretory glands surrounding the eye to increase corneal hydration.This would help to alleviate the symptoms associated with dry eyedisease, Sjögrens's syndrome is an autoimmune disease in which theimmune system attacks moisture-producing glands throughout the body,including the eye, mouth, skin, respiratory tissue, liver, vagina, andgut. Symptoms, include, dry eye, mouth, and vagina, as well as lungdisease. The disease is also associated with rheumatoid arthritis,systemic lupus, systemic sclerosis, and polymypositis/dermatomyositls.Defective protein trafficking is believed to cause the disease, forwhich treatment options are limited. Modulators of CFTR activity mayhydrate the various organs afflicted by the disease and help to elevatethe associated symptoms.

As discussed above, it is believed that the deletion of residue 508 inΔF508-CFTR prevents the nascent protein from folding correctly,resulting in the inability of this mutant protein to exit the ER, andtraffic to the plasma membrane. As a result insufficient amounts of themature protein are present at the plasma membrane and chloride transportwithin epithelial tissues is significantly reduced. In fact, thiscellular phenomenon of defective ER processing of ABC transporters bythe EE machinery, has been shown to be the underlying basis not only forcystic fibrosis disease, but for a wide range of other isolated andinherited diseases. The two ways that the ER machinery can malfunctionis either by loss of coupling to ER export of the proteins leading todegradation, or by the ER accumulation of these defective/misfoldedproteins (Aridor M, et al., Nature Med., 5(7), pp 745-751 (1999);Shastry, B. S., et al., Neurochem. International, 43, pp 1-7 (2003);Rutishauser, J., et al., Swiss Med Wkly, 132, pp 211-222 (2002);Morello, J P et al., TIPS, 21, pp. 466-469 (2000); Brass P., et al.,Human Mut., 14, pp. 186-198 (1999)). The diseases associated with thefirst class of ER malfunction are cystic fibrosis (due to misfoldedΔF508-CFTR as discussed above), hereditary emphysema (due toa1-antitrypsin; non Piz variants), hereditary hemochromatosis,coagulation-fibrinolysis deficiencies, such as protein C deficiency,Type 1 hereditary angioedema, lipid processing deficiencies, such asfamilial hypercholesterolemia, Type 1 chylomicronemia,abetalipoproteinemia, lysosomal storage diseases, such as I-celldisease/pseudo-Hurler, mucopolysaccharidoses (due to lysosomalprocessing enzymes), Sandhof/Tay-Sachs (due to β-hexosaminidase),Crigler-Hajjar type II (due to UDP-glucuronyl-sialyc-transferase),polyendocrinopathy/hyperinsulemia, diabetes mellitus (due to insulinreceptor), Laron dwarfism (due to growth hormone receptor),myleoperoxidase deficiency, primary hypoparathyroidism (due topreproparathyroid hormone), melanoma (due to tyrosinase). The diseasesassociated with the latter class of ER malfunction are glycanosis CDGtype 1, hereditary emphysema (due to α1-antitrypsin (PiZ variant),congenital hyperthyroidism, osteogenesis imperfecta (due to Type I, II,IV procollagen), hereditary hypofibrinogenemia (due to fibrinogen), ACTdeficiency (due to α1-antichymotrypsin), diabetes insipidus (DI),neurophyseal DI (due to vasopvessin hormone/V2-receptor), neprogenic DI(due to aquaporin II), Charcot-Marie Tooth syndrome (due to peripheralmyelin protein 22), Perlizaeus-Merzbacher disease, neurodegenerativediseases such as Alzheimer's disease (due to βAPP and presenilins),Parkinson's disease, amyotrophic lateral sclerosis, progressivesupranuclear plasy, Pick's disease, several polyglutamine neurologicaldisorders such as Huntington, spinocerebullar ataxia type I, spinal andbulbar muscular atrophy, dentatorubal pallidoluysian, and myotonicdystrophy, as well as spongiform encephalopathies, such as hereditaryCreutefeldt-Jakob disease (due to prion protein processing defect),Fabry disease (due to lysosomal α-galactosidase A), Straussler-Scheinkersyndrome, chronic obstructive pulmonary disease (COPD), dry eye disease,and Sjögren's Syndrome.

In addition to up-regulation of CFTR activity, reducing anion secretionby CFTR modulators may be beneficial for the treatment of secretorydiarrheas, in which epithelial water transport is dramatically increasedas a result of secretogogue activated chloride transport. The mechanisminvolves elevation of cAMP and stimulation of CFTR.

Although there are numerous causes of diarrhea, the major consequencesof diarrheal diseases, resulting from excessive chloride transport arecommon to all, and include dehydration, acidosis, impaired growth anddeath.

Acute and chronic diarrheas represent a major medical problem in manyareas of the world. Diarrhea is both a significant factor inmalnutrition and the leading cause of death (5,000,000 deaths/year) inchildren less than five years old.

Secretory diarrheas are also a dangerous condition in patients ofacquired immunodeficiency syndrome (AIDS) and chrome inflammatory boweldisease (IBD). Sixteen million travelers to developing countries fromindustrialized nations every year develop diarrhea, with the severityand number of cases of diarrhea varying depending on the country andarea of travel.

Diarrhea in barn animals and pets such as cows, pigs and horses, sheep,goats, cats and dogs, also known as scours, is a major cause of death inthese animals. Diarrhea can result from any major transition, such asweaning or physical movement, as well as in response to a variety ofbacterial or viral infections and generally occurs within the first fewhours of the animal's life.

The most common diarrheal causing bacteria is enterotoxigenic E-coli(ETEC) having the K99 pilus antigen. Common viral causes of diarrheainclude rotavirus and coronavirus. Other infectious agents includecryptosporidium, giardia lamblia, and salmonella, among others.

Symptoms of rotaviral infection include excretion of watery feces,dehydration and weakness. Coronovirus causes a more severe illness inthe newborn animals, and has a higher mortality rate than rotaviralinfection. Often, however, a young animal may be infected with more thanone virus or with a combination of viral and bacterial microorganisms atone time. This dramatically increases the severity of the disease.

Accordingly, there is a need for modulators of CFTR activity, andcompositions thereof, that can be used to modulate the activity of theCFTR in the cell membrane of a mammal.

There is a need for methods of treating CFTR-mediated diseases usingsuch modulators of CFTR activity.

There is a need for methods of modulating CFTR activity in an ex vivocell membrane of a mammal.

SUMMARY OF THE INVENTION

It has now been found that compounds of this invention, andpharmaceutically acceptable compositions thereof, are useful asmodulators of CFTR activity. These formula I or formula II:

or a pharmaceutically acceptable salt thereof, wherein R_(X), ring A, R,R^(AA), R^(BB), Z, and q are described below.

These compounds and pharmaceutically acceptable compositions are usefulfor treating or lessening the severity of a variety of diseases,disorders, or conditions, including, but not limits to, cystic fibrosis,hereditary emphysema, hereditary hemochromatosis,coagulation-fibrinolysis deficiencies, such as protein C deficiency,Type 1 hereditary angioedema, lipid processing deficiencies, such asfamilial hypercholesterolemia, Type 1 chylomicronemia,abetalipoprotememia, lysosomal storage diseases, such as I-celldisease/pseudo-Hurler, mucopolysaccharidoses, Sandhof/Tay-Sachs,Crigler-Najjar type II, polyendocrinopathy/hyperinsulemia, diabetesmellitus, laron dwarfism, myleoperoxidase deficiency, primaryhypoparathyroidism, melanoma, glycanosis COG type 1, hereditaryemphysema, congenital hyperthyroidism, osteogenesis imperfecta,hereditary hypofibrinogenemia, ACT deficiency, diabetes insipidus (di),neurophyseal di, neprogenic DI, Charcot-Marie Tooth syndrome,Perlizaeus-Merzbacher disease, neurodegenerative diseases such asAlzheimer's disease. Parkinson's disease, amyotrophic lateral sclerosis,progressive supranuclear plasy, Pick's disease, several polyglutamineneurological disorders asuch as Huntington, spinocerebellar ataxia typeI, spinal and bulbar muscular atrophy, dentatorubal pallidoluysian, andmyotonic dystrophy, as well as spongiform encephalopathies, such ashereditary Creutefeldt-Jakob disease, Fabry disease,Straussler-Scheinker syndrome, COPD, dry-eye disease, and Sjogren'sdisease.

DETAILED DESCRIPTION OF THE INVENTION I. General Description ofCompounds of the Invention

The present invention provides compounds of formula I or formula II:

or a pharmaceutically acceptable salt thereof, wherein:

each R^(X) is independently hydrogen, halo, CF₃, C1-C4 alkyl, or —OC1-C4alkyl; provided that both R^(X) are not simultaneously hydrogen; or thetwo R^(X), taken together form ring (a):

X is CH₂, CF₂, CH₂—CH₂, or CF₂—CF₂;

ring A is 3-7 membered monocyclic cycloalkyl ring;

R^(AA) and R^(BB) taken together with the nitrogen atom, form apyrrolidinyl ring substituted with OR′; R′ is hydrogen or C1-C6aliphatic, wherein up to two carbon units of said aliphatic areoptionally and independently replaced by —CO—, —CS—, —COCO—, —CONR—,—CONRNR—, —CO₂—, —OCO—, —NRCO₂—, —O—, —NRCONR—, —OCONR—, —NRNR,—NRNRCO—, —NRCO—, —S—, —SO, —SO₂—, —NR—, —SO₂NR—, NRSO₂—, or —NRSO₂NR—;

R is hydrogen or C1-C6 aliphatic;

Z is an electron withdrawing substituent; and

q is 0-3.

As used herein, the following definitions shall apply unless otherwiseindicated.

The term “CFTR” as used herein means cystic fibrosis transmembraneconductance regulator or a mutation thereof capable of regulatoractivity, including, but not limited to, ΔF508 CFTR and G55ID CFTR (see,e.g., http://www.genet.sickkids.on.ca/cftr/, for CFTR mutations).

The term “modulating” as used herein means increasing or decreasing by ameasurable amount.

The term “correction” as used herein means increasing the number of CFTRin a membrane of a cell.

The term “potentiator” as used herein means a compound that increasesthe gating activity of CFTR in a membrane of a cell.

The term “electron withdrawing substituent”, as used herein means anatom or a group that is electronegative relative to hydrogen. See, e.g.,“Advanced Organic Chemistry: Reactions, Mechanisms, and Structure,”Jerry March, 4^(th) Ed., John Wiley & Sons (1992), e.g., pp. 14-16,18-19, etc. Exemplary such substituents include halo, CN, COOH, CF₃,etc.

For purposes of this invention, the chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 75^(th) Ed. Additionally, generalprinciples of organic chemistry are described in “Organic Chemistry”,Thomas Sorrell, University Science Books, Sausalito: 1999, and “March'sAdvanced Organic Chemistry”, 5^(th) Ed., Ed.: Smith, M. B. and March,J., John Wiley & Sons, New York: 2001, the entire contents of which arehereby incorporated by reference.

Unless otherwise specified, the term “aliphatic” or “aliphatic group” byitself, as used herein, means a straight-chain (i.e., unbranched) orbranched, substituted or unsubstituted hydrocarbon chain that iscompletely saturated or that contains one or more units of unsaturation,or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completelysaturated or that contains one or more units of unsaturation, but whichis not aromatic (also referred to herein as “carbocycle,”“cycloaliphatic” or “cycloalkyl”), that has a single point of attachmentto the rest of the molecule. Unless otherwise specified, aliphaticgroups contain 1-20 aliphatic carbon atoms. In some embodiments,aliphatic groups contain 1-10 aliphatic carbon atoms. In otherembodiments, aliphatic groups contain 1-5 aliphatic carbon atoms. Instill other embodiments, aliphatic groups contain 1-6 aliphatic carbonatoms, and in yet other embodiments aliphatic groups contain 1-4aliphatic carbon atoms. In some embodiments, “cycloaliphatic” (or“carbocycle” or “cycloalkyl”) refers to a monocyclic C3-C8 hydrocarbonor bicyclic C8-C12 hydrocarbon that is completely saturated or thatcontains one or more units of unsaturation, but which is not aromatic,that has a single point of attachment to the rest of the moleculewherein any individual ring in said bicyclic ring system has 3-7members. Suitable aliphatic groups include, but are not limited to,linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynylgroups and hybrids thereof such as (cycloalkyl)alkyl,(cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

The term “unsaturated”, as used herein, means that a moiety has one ormore units of unsaturation.

Unless stereochemically specified, structures depicted herein are alsomeant to include all isomeric (e.g., enantiomeric, diastereomeric, andgeometric (or conformational)) forms of the structure; for example, theR and S configurations for each asymmetric center, (Z) and (E) doublebond isomers, and (Z) and (E) conformational isomers, including mixturescontaining an excess of one enantiomer relative to the other enantiomeror an excess of one diastereomer relative to another. Unless otherwisespecified, single stereochemical isomers as well as enantiomeric,diastereomeric, and geometric (or conformational) mixtures of thepresent compounds are within the scope of the invention. Unlessotherwise stated, all tautomeric forms of the compounds of the inventionare within the scope of the invention. Additionally, unless otherwisestated, structures depicted herein are also meant to include compoundsthat differ only in the presence of one or more isotopically enrichedatoms. For example, compounds having the present structures except forthe replacement of hydrogen by deuterium or tritium, or the replacementof a carbon by a ¹³C- or ¹⁴C-enriched carbon are within the scope ofthis invention. Such compounds are useful, for example, as analyticaltools or probes in biological assays.

In one embodiment, each R^(X) is independently hydrogen, halo, or CF₃;provided that both R^(X) are not simultaneously hydrogen. In anotherembodiment, one R^(X) is hydrogen and the other R^(X) is halo or CF₃. Inanother embodiment, both R^(X) are halo.

In one embodiment, the two R^(X) groups taken together form ring (a).

In certain embodiments, X is Ch₂. In other embodiments, X is CF₂. Or, Xis CH₂—CH₂. In certain other embodiments, X is CF₂—CF₂;

In one embodiment, ring A is cyclopropyl, cyclopentyl, or cyclohexyl. Inanother embodiment, ring A is cyclopropyl or cyclopentyl. In certainembodiments, ring A is cyclopropyl.

In one embodiment, R is hydrogen. Or, R is C1-C6 alkyl. Exemplary Rincludes methyl, ethyl, or propyl.

In one embodiment, R is hydrogen. Or, R′ is C1-C6 alkyl. Exemplary R′include methyl, ethyl, propyl, or C(O)Me.

In one embodiment, R^(AA) and R^(BB), taken together, form apyrrolidinyl with an OH substituent.

In one embodiment, Z is selected from halo, CF₃, ordifluoromethylenedioxy. In another embodiment, Z is fluoro or chloro.

In one embodiment, q is 0. Or, q is 1-2. In certain embodiments, q is 1.Or, q is 2.

In another embodiment, compounds of formula I or formula II comprise oneor more, and preferably all, of the following features:

-   -   the two R^(X) are taken together to form ring (a);    -   X is CH₂;    -   ring A is cyclopropyl;    -   R′ is hydrogen;    -   q is 1 or 2; and    -   Z is halo, CF₃, or difluoromethylenedioxy.

In another embodiment, compounds of formula I or formula II comprise oneor more, and preferably all, of the following features:

-   -   the two R^(X) are taken together to form ring (a);    -   X is CF₂;    -   R is hydrogen;    -   ring A is cyclopropyl;    -   R′ is hydrogen;    -   q is 1 or 2; and    -   Z is halo, CF₃, or difluoromethylenedioxy.

In another embodiment, compounds of formula I or formula II comprise oneor more, and preferably all, of the following features:

-   -   the two R^(X) are taken together to form ring (a);    -   X is CF₂;    -   ring A is cyclopropyl;    -   R′ is hydrogen;    -   q is 1 or 2; and    -   Z is halo, CF₃, or difluoromethylenedioxy.

In another embodiment, compounds of formula I or formula II comprise oneor more, and preferably all, of the following features:

-   -   the two R^(X) are taken together to form ring (a);    -   X is CF₂;    -   R is hydrogen;    -   ring A is cyclopropyl;    -   R′ is hydrogen;    -   q is 1 or 2; and

Z is halo, CF₃, or difluoromethylenedioxy. In one embodiment ofcompounds of formula I, R^(AA) and R^(BB), taken together with thenitrogen atom, form the following ring (i):

In one embodiment of compounds of formula II, R^(AA) and R^(BB), takentogether with the nitrogen atom, form the following ring (ii):

In one embodiment of compounds of formula I, R^(AA) and R^(BB), takentogether with the nitrogen atom, form the following ring (iii):

In one embodiment of compounds of formula II, R^(AA) and R^(BB), takentogether with the nitrogen atom, form the following ring (iv):

In an alternative embodiment, the present invention providesintermediates having formula I′ or formula II′:

wherein:

-   -   R^(X), R, ring A, Z, and q are as defined above;    -   L is a linker selected from C(O), SO₂;    -   is 0 or 1; and    -   CA is a suitable chiral auxiliary.

The term “chiral auxiliary” as used herein means an asymmetric moleculeor molecular fragment that is used to achieve the chemical resolution ofa racemic or diastereomeric mixture. Such chiral auxiliaries may possessone chiral center such as methylbenylamine or several chiral centerssuch as menthol. The purpose of a chiral auxiliary, once built into thestarting material, is to allow simple separation of the resultingdiastereomeric mixture. See, for example, J. Jacques et al.,“Enantiomers, Racemates And Resolutions,” pp. 251-369, John Wiley &Sons, New York (1981); E. L. Eliel & S. H. Wilen, “Stereochemistry ofOrganic Compounds.” pp. 868-870, John Wiley & Sons (1994).

Suitable chiral auxiliaries useful in the present invention includethose that are amenable to attachment to the linker L above (i.e., pis 1) or directly to the oxygen atom (i.e., p is 0). Exemplary suchchiral auxiliaries are found in, e.g., E. L. Eliel & S. H. Wilen, ibid,pp. 337-340.

In one embodiment, CA, L, p, and the oxygen atom linked thereto, takentogether, is (+)-10-camphorsulfonate, (1S, 4R)-9−)-ω-camphanicester,(1R, 2S, 5R)-(−) mentholcarbonate, (1S, 2R,5S)-(+)-mentholcarbonate, (1R, 2R)-1-phenyl-2-cyclopropylester, or(3R)-tetrahydrofuran-3-carbonate.

Exemplary compounds of the present invention are shown below in Table 1.

TABLE 1

1

2

3

4

5

6

7

8

9

10

11

12

13

In one embodiment, the present invention provides a method for producinga compound of formula I or formula II:

comprising the step of reacting under a first suitable conditions acompound of formula R-1 with either a compound of formula I-A to producesaid compound of formula I, or a compound of formula II-A to producesaid compound of formula II:

-   -   wherein:        each R^(X) is independently hydrogen, halo, CF₃, C1-C4 alkyl, or        —OC1-C4 alkyl; provided that both R^(X) are not simultaneously        hydrogen; or    -   the two R^(X), taken together form ring (a):

X is CH₂, CF₂, CH₂—CH₂, or CF₂—CF₂;

-   -   ring A is 3-7 membered monocyclic cycloalkyl ring;    -   R^(AA) and R^(BB), taken together with the nitrogen atom, form a        pyrrolidmyl ring substituted with OR′;    -   R′ is hydrogen or C1-C6 aliphatic, wherein up to two carbon,        units of said aliphatic are optionally and independently        replaced by —CO—, —CS—, —COCO—, —CONR—, —CONRNR—, —CO₂—, —OCO—,        —NRCO₂—, —O—, —NRCONR—, —OCONR—, —NRNR, —NRNRCO—, —NRCO—, —S—,        -SO, —SO, —SO₂—, —NR—, —SO₂NR—, NRSO₂—, or —NRSO₂NR—;    -   R is hydrogen or C1-C6 aliphatic;    -   Z is an electron withdrawing substituent; and    -   q is 0-3; and    -   LG₁ is a first, suitable leaving group

As used herein, the term “first suitable conditions” means conditionssuitable to effectuate the reaction between compound of formula I-A andcompound of formula R-1 or between compound of formula II-A and compoundof formula R-1. Such suitable conditions include, e.g., a first suitablesolvent, a first suitable temperature, and a suitable reducing agent.One of skill in the art will be well aware of various such suitableconditions that effectuate the reaction between compound of formula I-Aand compound of formula R-1 or between compound of formula II-A andcompound of Formula R-1.

In one embodiment, the first suitable solvent is a polar aprotic, apolar protic solvent, an apolar solvent, or a suitable combinationthereof. Exemplary solvents useful as first suitable solvent includemethanol, ethanol, propanol, isopropanol, t-butanol, dichloromethane,dichloroethane, toluene, tetrahydrofuran, dioxane, diethylether,dimethylether, acetonitrile, dimethylformamide, DMAC, or NMP.

In one embodiment, the first suitable temperature is a temperature thatis sufficient to effectuate the reaction between compound of formula I-Aand compound of formula R-1 in the first suitable solvent. In anotherembodiment, the first suitable temperature is a temperature that issufficient to effectuate the reaction between compound of formula II-Aand compound of formula R-1 in the first suitable solvent. Exemplaryfirst suitable temperature includes between about 0° C. to about 110° C.In one embodiment, the first suitable temperature is between about 0° C.to about 25° C.

In one embodiment, the suitable reducing agent is a reducing agent thatis capable effectuating the reaction between compound of formula I-A andcompound of formula R-1. In another embodiment, the suitable reducingagent is a reducing agent that is capable effectuating the reactionbetween compound of formula II-A and compound of formula R-1. One ofskill in the art will be well aware of suitable reducing agents for thatreaction. Agents suitable for the present invention include ametallo-borohydride or a reagent capable of catalytic Hydrogenation.Exemplary such suitable reducing agents include sodium borohydride,sodium cyanoborohydride, lithium borohydride, sodiumtriacetoxyborohydride, calcium borohydride, hydrogene in the presence ofa suitable metal catalyst such as Pd/C.

In another embodiment, LG₁ is a first suitable leaving group that iscapable of displacement to produce compound of formula I, or compound offormula II. See, “Advanced Organic Chemistry: Reactions, Mechanisms, andStructure,” pp. 339-357, Jerry March, 4^(th) Ed., John Wiley & Sons(1992).

The method according to claim 1, wherein LG₁ is selected fromalkysulfonate, arylsulfonate, halide, alkyl carboxylate.

In one embodiment, the compound of formula I-A is produced from formulaI-B:

-   -   wherein [CA] is a suitable chiral auxiliary;    -   said method comprising the step of removing said chiral auxilary        under a second suitable conditions.

In an alternative embodiment, the compound of formula II-A is producedfrom formula II-B.

-   -   wherein [CA] is a suitable chiral auxiliary;    -   said method comprising the step of removing said chiral auxilary        under a second suitable conditions.

A “suitable chiral auxiliary” above in compound of formula II-B orformula I-B is a chiral auxiliary that is capable of attachment to anamino group. An isomeric mixture of a compound containing such an aminogroup with a suitable chiral auxiliary attached thereto is readilyseparated into its individual isomers by suitable separation means. See.for example, J. Jacques et al., “Enantiomers, Racemates AndResolutions,” pp. 251-369, John Wiley & Sons, New York (1981); E. L.Eliel & S. H. Wilen, “Sterochemistry of Organic Compounds,” pp. 868-870,John Wiley & Sons (1994).

In one embodiment, said suitable chiral auxiliary is an alkylsulfoxylgroup.

In another embodiment, said suitable second conditions comprises asuitable protic acid and a suitable second solvent.

In one embodiment, said suitable second solvent is selected from a polaraprotic solvent or a protic solvent. Exemplary polar aprotic solventsinclude dioxane, tetrahydrofurane, diethyl ether, dichloromethane, etc.Exemplary protic solvents include methanol, ethanol, i-propanaol,t-butanol, etc.

In one embodiment, said suitable second solvent is a polar aproticsolvent.

In another embodiment, said compound of formula I-B and compound offormula II-B is produced by reacting, respectively, a compound offormula I-C or formula II-C with a compound of formula R-2 under a thirdsuitable conditions:

-   -   wherein:    -   R is hydrogen or C1-C6 aliphatic;    -   R¹ is hydrogen or a first suitable protecting group;    -   [CA] is a suitable chiral auxiliary; and    -   LG₂ is a second suitable leaving group.

In one embodiment, LG₂ is selected from halide, OC(O)(C1-C6 alkyl),pentafluorophenoxy, C1-C6 alkoxy, OCO₂(C1-C6 alkyl) or hydroxy.

In one embodiment, R¹ is hydrogen.

In another embodiment, said third suitable conditions comprises a thirdsuitable coupling agent and a third suitable solvent.

In another embodiment, said suitable coupling agent is selected fromDCC, DCI, HATU, TCPH, or HBTU.

In one embodiment, said third suitable solvent is selected fromdichloromethane, dioxane, acetonitrile, DMF, dichloroethane, ortetrahydrofuran.

In another embodiment, said compound of formula I-C or formula II-C isproduced from an isomeric mixture of a compound of formula R-3;

-   -   wherein;    -   R, Z, and q are as defined above;    -   PG₁ s a second suitable protecting group; and    -   [CA] is a suitable chiral auxiliary;    -   said method comprising two steps, wherein one of said two steps        is separating said isomeric mixture using suitable separation        means, and the other of said two steps is the conversion of PG₁        to R¹ in a fourth suitable conditions.

In one embodiment, said suitable separation means comprises suitablechromatographic means. Exemplary such means include columnchromatography or thin layer chromatography.

In another embodiment, said suitable separation means comprises suitablecrystallization means.

In another embodiment, said fourth suitable conditions comprises asuitable deproiectlng reagent and a fourth suitable solvent. Anexemplary suitable deprotectnig reagent is trifluoroacetic acid.

In one embodiment, said fourth suitable solvent is a polar aptoticsolvent. Exemplary solvents include dichloromethane, tetrahydrofuran,dioxane, diethyl ether, etc.

In another embodiment, said compound of formula R-3 is produced from acompound of formula R-4 and a compound of formula R-5;

-   -   wherein:        -   PG₂ is a second suitable protecting group;        -   M is a suitable metal cation; and        -   [CA] is a suitable chiral auxiliary;    -   said method comprising the steps of reacting said compound of        formula R-4 with said compound of formula a R-5 in a fifth        suitable conditions.

In one embodiment, said M is selected from Li³⁰ , Na⁺, or Mg⁺⁺.

In another embodiment, said PG₁ is selected from an alkylcarbamate,trifluoroacetyl, trialkylsilyl, or pivaloyl. Or, said PG₁ is BOC ortrimethylsilyl.

In one embodiment, said fifth suitable conditions comprises a fifthsuitable solvent and a fifth suitable temperature. In one embodiment,said suitable temperature is about −78 degrees C.

In another embodiment, said fifth suitable solvent is tetrahydrofuran.

In an alternative embodiment, said compound of formula R-1 is:

Or, said compound of formula R-1 is:

In another embodiment, said compound of formula I or formula II areselected from Table 1.

4. General Synthetic Schemes

The compounds of this invention may be prepared by methods known in theart. Exemplary synthetic routes to prepare compounds of this inventionare illustrated below.

Scheme I-A below illustrates a process for producing intermediate A:

-   -   a) 50% NaOH, Y(CH₂)nY, BTEAC (benzyl triethyl ammonium        chloride); Y=suitable leaving group.

Scheme I-B below illustrates a process for producing intermediate B.

Scheme I-C below illustrates a process for producing compounds offormula I or formula II:

Scheme I-D illustrates the synthesis of an exemplary compound of thepresent invention.

Scheme II-A below illustrates another exemplary process for preparingcompounds of the present invention using a chiral auxiliary.

Scheme II-B below illustrates a process for preparing an exemplarycompound of the present invention using a chiral auxiliary.

Scheme III below illustrates another process for preparing a compound ofthe present invention.

Further illustrative examples for preparing compounds of the presentinvention are recited below.

5. Uses, Formulation and Administration

Pharmaceutically Acceptable Compositions

As discussed above, the present invention provides compounds that areuseful as modulators of CFTR and thus are useful in the treatment ofdisease, disorders or conditions such as Cystic fibrosis, Hereditaryemphysema, Hereditary hemochromatosis, Coagulation-Fibrinolysisdeficiencies, such as Protein C deficiency. Type 1 hereditaryangioedema, Lipid processing deficiencies, such as Familialhypercholesterolemia, Type 1 chylomlcronemia, Abetalipoproteioemia,Lysosomal storage diseases, such as I-cell disease/Pseudo-Hurler,Mucopolysaccharidoses, Sandhof/Tay-Sachs, Crigler-Najjar type II,Polyendocrinopathy/Hyperinsulemia, Diabetes mellitus, Laron dwarfism,Myleoperoxidase deficiency, Primary hypoparathyroidism, Melanoma,Glycanosis CDG type 1, Hereditary emphysema, Congenital hyperthyroidism,Osteogenesis imperfecta, Hereditary hypofibrinogenemia, ACT deficiency;Diabetes insipidus (DI), Neorophyseal DI, Neprogenic DI, Charcot-MarieTooth syndrome, Perlizaeus-Merzbacher disease, neurodegenerativediseases such as Alzheimer's disease, Parkinson's disease, Amyotrophiclateral sclerosis, Progressive supranuclear plasy, Pick's disease,several polyglutamlne neurological disorders asuch as Huntington,Spinocerebullar ataxia type I, Spinal and bulbar muscular atrophy,Dentatorubal pallidoloysian, and Myotonic dystrophy, as well asSpongiform encephalopathies, such as Hereditary Creutzfeldt-Jakobdisease (due to Prion protein processing defect), Fabry disease andStraussler-Scheinker syndrome.

Accordingly, in another aspect of the present invention,pharmaceutically acceptable compositions are provided, wherein thesecompositions comprise any of the compounds as described herein, andoptionally comprise a pharmaceutically acceptable carrier, adjuvant orvehicle. In certain embodiments, these compositions optionally furthercomprise one or more additional therapeutic agents.

It will also be appreciated that certain of the compounds of presentinvention can exist in free form for treatment, or where appropriate, asa pharmaceutically acceptable derivative thereof. According to thepresent invention, a pharmaceutically acceptable derivative includes,but is not limited to, pharmaceutically acceptable salts, esters, saltsof such esters, or any other adduct or derivative which uponadministration to a patient in need is capable of providing, directly orindirectly, a compound as otherwise described herein, or a metabolite orresidue thereof.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgement,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. A“pharmaceutically acceptable salt” means any non-toxic salt or salt ofan ester of a compound of this invention that, upon administration to arecipient, is capable of providing, either directly or indirectly, acompound of this invention or an inhibitorily active metabolite orresidue thereof. As used herein, the term “inhibitorily activemetabolite or residue thereof” means that a metabolite or residuethereof is also an inhibitor of an ATP-Binding Cassette Transporters.

Pharmaceutically acceptable salts are well known in the art. Forexample, S. M. Berge, et al. describe pharmaceutically acceptable saltsin detail in J. Pharmaceutical Sciences, 1911, 66, 1-19, incorporatedherein by reference. Pharmaceutically acceptable salts of the compoundsof this invention include those derived from suitable inorganic andorganic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. This inventionalso envisions the quaternization of any basic nitrogen-containinggroups of the compounds disclosed herein. Water or oil-soluble ordispersable products may be obtained by such quaternization.Representative alkali or alkaline earth metal salts include sodium,lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, loweralkyl sulfonate and aryl sulfonate.

As described above, the pharmaceutically acceptable compositions of thepresent invention additionally comprise a pharmaceutically acceptablecarrier, adjuvant, or vehicle, which, as used herein, includes any andall solvents, diluents, or other liquid vehicle, dispersion orsuspension aids, surface active agents, isotonic agents, thickening oremulsifying agents, preservatives, solid binders, lubricants and thelike, as suited to the particular dosage form desired. Remington'sPharmaceutical Sciences, Sixteenth Edition, E. W. Martin (MackPublishing Co., Easton, Pa., 1980) discloses various carriers used informulating pharmaceutically acceptable compositions and knowntechniques for the preparation thereof. Except insofar as anyconventional carrier medium is incompatible with the compounds of theinvention, such as by producing any undesirable biological effect orotherwise interacting in a deleterious manner with any othercomponent(s) of the pharmaceutically acceptable composition, its use iscontemplated to be within the scope of this invention. Some examples ofmaterials which can serve as pharmaceutically acceptable carriersinclude, but are not limited to, ion exchangers, alumina, aluminumstearate, lecithin, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, or potassiumsorbate, partial glyceride mixtures of saturated vegetable fatty acids,water, salts or electrolytes, such as protamine sulfate, disodiumhydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zincsalts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, woolfat, sugars such as lactose, glucose and sucrose; starches such as comstarch and potato starch; cellulose and its derivatives such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; powderedtrigacanth; malt; gelatin; talc; excipients such as cocoa butter andsuppository waxes; oils such as peanut oil, cottonseed oil; sunfloweroil; sesame oil; olive oil; corn oil and soybean oil; glycols; such apropylene glycol or polyethylene glycol; esters such as ethyl oleate andethyl laurate; agar; buffering agents such as magnesium hydroxide andaluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline;Ringer's solution; ethyl alcohol, and phosphate buffer solutions, aswell as other non-toxic compatible lubricants such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releasingagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator.

Uses of Compounds and Pharmaceutically Acceptable Compositions

In yet another aspect, the present invention provides a method oftreating a condition disease, or disorder implicated by CFTR. In certainembodiments, the present invention provides a method of treating acondition, disease, or disorder implicated by a deficiency of CFTR, themethod comprising administering a composition comprising a compound offormula (I) to a subject, preferably a mammal, in need thereof.

In certain preferred embodiments, the present invention provides amethod of treating cystic fibrosis, hereditary emphysema (due toa1-antitrypsin; non Piz variants), hereditary hemochromatosis,coagulation-fibrinolysis deficiencies, such as Protein C deficiency,Type I hereditary angioedema, lipid processing deficiencies, such asfamilial hypercholesterolemia, Type Type I chylomicronemia,abetalipoproteinemia, lysosomal storage diseases, such as I-celldisease/pseudo-Hurler, mucopolysaccharidoses (due to lysosomalprocessing enzymes), Sandhof/Tay-Sachs (due to β-hexosaminidase),Crigler-Najjar type II (due to UDP-glucorouyl-sialyc-transfease),polyendocrinopathy/hyperinsulemia, diabetes mellitus (due to insulinreceptor), Laron dwarfism (due to growth hormone receptor),myleoperoxidase deficiency, primary hypoparathyroidism (due topreproparathyroid hormone), melanoma (due to tyrosinase). The diseasesassociated with the latter class of ER malfunction are glycanosis CDGtype 1, hereditary emphysema (due to α1-antitrypsin (PiZ variant),congenital hyperthyroidism, osteogenesis imperfecta (due to Type I, II,IV procollagen), hereditary hypofibrinogenemia (due to fibrinogen), ACTdeficiency (due to α1-antichymotrypsin), diabetes insipidus (DI),neorophyseal DI (due to vasopvessin hormone/V2-receptor, neprogenic DI(due to aquaporin II), Charcot-Marie Tooth syndrome (due to peripheralmyelin protein 22), Perlizaeus-Merzbacher disease, neurodegenerativediseases such as Alzheimer's disease (due to βAPP and presenilins),Parkinson's disease, amyotrophic lateral sclerosis, progressivesupranuclear plasy, Pick's disease, several polyglutamine neurologicaldisorders such as Huntington, spinocerebullar ataxia type 1, spinal andbulbar muscular atrophy, dentatorubal pallidoluysian, and myotonicdystrophy, as well as spongiform encephalopathies, such as hereditaryCreutzfeldt-Jakob disease (due to prion protein processing defect),Fabry disease (due to lysosomal α-galactosidase A), Straussler-Scheinkersyndrome, chronic obstructive pulmonary disease (COPD), dry eye disease,and Sjögren's Syndrome, comprising the step of administering to saidmammal an effective amount of a composition comprising a compound offormula (I), or a preferred embodiment thereof as set forth above.

According to an alternative preferred embodiment, the present inventionprovides a method of treating cystic fibrosis comprising the step ofadministering to said mammal a composition comprising the step ofadministering to said mammal an effective amount of a compositioncomprising a compound of formula (I), or a preferred embodiment thereofas set forth above.

According to the invention an “effective amount” of the compound orpharmaceutically acceptable composition is that amount effective fortreating or lessening the severity of one or more of cystic fibrosis,hereditary emphysema (due to a1-antitrypsin; non Piz variants),hereditary hemochromatosis, coagulation-fibrinolysis deficiencies, suchas Protein C deficiency, Type I hereditary angioedema, lipid processingdeficiencies, such as familial hypercholesterolemia, Type Ichylomicronemia, abetalipoproteinemia, lysosomal storage diseases, suchas I-cell disease/pseudo-Hurler, mucopolysaccharidoses (due to lysosomalprocessing enzymes), Sandhof/Tay-Sachs (due to β-hexosaminidase),Crigler-Najjar type II (due to UDP-glucorouyl-sialyc-transfease),polyendocrinopathy/hyperinsulemia, diabetes mellitus (due to insulinreceptor), Laron dwarfism (due to growth hormone receptor),myleoperoxidase deficiency, primary hypoparathyroidism (due topreproparathyroid hormone), melanoma (due to tyrosinase). The diseasesassociated with the latter class of ER malfunction are glycanosis CDGtype 1, hereditary emphysema (due to α1-antitrypsin (PiZ variant),congenital hyperthyroidism, osteogenesis imperfecta (due to Type I, II,IV procollagen), hereditary hypofibrinogenemia (due to fibrinogen), ACTdeficiency (due to α1-antichymotrypsin), diabetes insipidus (DI),neorophyseal DI (due to vasopvessin hormone/V2-receptor, neprogenic DI(due to aquaporin II), Charcot-Marie Tooth syndrome (due to peripheralmyelin protein 22), Perlizaeus-Merzbacher disease, neurodegenerativediseases such as Alzheimer's disease (due to βAPP and presenilins),Parkinson's disease, amyotrophic lateral sclerosis, progressivesupranuclear plasy, Pick's disease, several polyglutamine neurologicaldisorders such as Huntington, spinocerebullar ataxia type 1, spinal andbulbar muscular atrophy, dentatorubal pallidoluysian, and myotonicdystrophy, as well as spongiform encephalopathies, such as hereditaryCreutzfeldt-Jakob disease (due to prion protein processing defect),Fabry disease (due to lysosomal α-galactosidase A), Straussler-Scheinkersyndrome, chronic obstructive pulmonary disease (COPD), dry eye disease,and Sjögren's Syndrome,.

The compounds and compositions, according to the method of the presentinvention, may be administered using any amount and any route ofadministration effective for treating or lessening the severity of oneor more of cystic fibrosis, hereditary emphysema (due to a1-antitrypsin;non Piz variants), hereditary hemochromatosis, coagulation-fibrinolysisdeficiencies, such as Protein C deficiency, Type Ihereditary angioedema,lipid processing deficiencies, such as familial hypercholesterolemia,Type I chylomicronemia, abetalipoproteinemia, lysosomal storagediseases, such as I-cell disease/pseudo-Hurler, mucopolysaccharidoses(due to lysosomal processing enzymes), Sandhof/Tay-Sachs (due toβ-hexosaminidase), Crigler-Najjar type II (due toUDP-glucorouyl-sialyc-transfease), polyendocrinopathy/hyperinsulemia,diabetes mellitus (due to insulin receptor), Laron dwarfism (due togrowth hormone receptor), myleoperoxidase deficiency, primaryhypoparathyroidism (due to preproparathyroid hormone), melanoma (due totyrosinase). The diseases associated with the latter class of ERmalfunction are glycanosis CDG type 1, hereditary emphysema (due toα1-antitrypsin (PiZ variant), congenital hyperthyroidism, osteogenesisimperfecta (due to Type I, II, IV procollagen), hereditaryhypofibrinogenemia (due to fibrinogen), ACT deficiency (due toα1-antichymotrypsin), diabetes insipidus (DI), neorophyseal DI (due tovasopvessin hormone/V2-receptor, neprogenic DI (due to aquaporin II),Charcot-Marie Tooth syndrome (due to peripheral myelin protein 22),Perlizaeus-Merzbacher disease, neurodegenerative diseases such asAlzheimer's disease (due to βAPP and presenilins), Parkinson's disease,amyotrophic lateral sclerosis, progressive supranuclear plasy, Pick'sdisease, several polyglutamine neurological disorders such asHuntington, spinocerebullar ataxia type I, spinal and bulbar muscularatrophy, dentatorubal pallidoluysian, and myotonic dystrophy, as well asspongiform encephalopathies, such as hereditary Creutzfeldt-Jakobdisease (due to prion protein processing defect), Fabry disease (due tolysosomal α-galactosidase A), Straussler-Scheinker syndrome, chronicobstructive pulmonary disease (COPD), dry eye disease, and Sjögren'sSyndrome,

The exact amount required will vary from subject to subject, dependingon the species, age, and general condition of the subject, the severityof the infection, the particular agent, its mode of administration, andthe like. The compounds of the invention are preferably formulated indosage unit form for ease of administration and uniformity of dosage.The expression “dosage unit form” as used herein refers to a physicallydiscrete unit of agent appropriate for the patient to be treated. Itwill be understood, however, that the total daily usage of the compoundsand compositions of the present invention will be decided by theattending physician within the scope of sound medical judgment. Thespecific effective dose level for any particular patient or organismwill depend upon a variety of factors including the disorder beingtreated and the severity of the disorder; the activity of the specificcompound employed; the specific composition employed; the age, bodyweight, general health, sex and diet of the patient; the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed, andlike factors well known in the medical arts. The term “patient”, as usedherein, means an animal, preferably a mammal, and most preferably ahuman.

The pharmaceutically acceptable compositions of this invention can beadministered to humans and other animals orally, rectally, parenterally,intracisternally, intravaginally, intraperitoneally, topically (as bypowders, ointments, or drops), bucally, as an oral or nasal spray, orthe like, depending on the severity of the infection being treated. Incertain embodiments, the compounds of the invention maybe administeredorally or parenterally at dosage levels of about 0.01 mg/kg to about 50mg/kg and preferably from about 1 mg/kg to about 25 mg/kg, of subjectbody weight per day, one or more times a day, to obtain the desiredtherapeutic effect.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters, of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenteraily acceptablediluent or solvent, for example, as a solution, in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium, chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a compound of the present invention,it is often desirable to slow the absorption of the compound fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the compound thendepends upon its rate of dissolution that, in turn, may depend uponcrystal size and crystalline form. Alternatively, delayed absorption ofa parenterally administered compound form is accomplished by dissolvingor suspending the compound in an oil vehicle. Injectable depot forms aremade by forming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) distintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption acclerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polethylene glycols and the like.

The active compounds can also be in microencapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed, under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, eardrops, and eye drops are also contemplated asbeing within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms are prepared by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

As described generally above, the compounds of the invention are usefulas modulators of CFTR. Thus, without wishing to be bound by anyparticular theory, the compounds and compositions are particularlyuseful for treating or lessening the severity of a disease, condition,or disorder where hyperactivity or inactivity of CFTR is implicated inthe disease, condition, or disorder. When hyperactivity or inactivity ofan CFTR is implicated in a particular disease, condition, or disorder,the disease, condition, or disorder may also be referred to as a“CFTR-mediated disease, condition or disorder”. Accordingly, in anotheraspect, the present invention provides a method for treating orlessening the severity of a disease, condition, or disorder wherehyperactivity or inactivity of an CFTR is implicated in the diseasestate.

The activity of a compound utilized in this invention as a modulator ofan CFTR may be assayed according to methods described generally in theart and in the Examples herein.

It will also be appreciated that the compounds and pharmaceuticallyacceptable compositions of the present invention can be employed Incombination therapies, that is, the compounds and pharmaceuticallyacceptable compositions can be administered concurrently with, prior to,or subsequent to, one or more other desired therapeutics or medicalprocedures. The particular combination of therapies (therapeutics orprocedures) to employ in a combination regimen will take into accountcompatibility of the desired therapeutics and/or procedures and thedesired therapeutic effect to be achieved. It will also be appreciatedthat the therapies employed may achieve a desired effect for the samedisorder (for example, an inventive compound may be administeredconcurrently with another agent used to treat the same disorder), orthey may achieve different effects (e.g., control of any adverseeffects). As used herein, additional therapeutic agents that arenormally administered to treat or prevent a particular disease, orcondition, are known as “appropriate for the disease, or condition,being treated”.

The amount of additional therapeutic agent present in the compositionsof this invention will be no more than the amount that would normally beadministered In a composition comprising that therapeutic agent as theonly active agent. Preferably the amount of additional therapeutic agentin the presently disclosed compositions will range from about 50% to100% of the amount normally present in a composition comprising thatagent as the only therapeutically active agent.

The compounds of this invention or pharmaceutically acceptablecompositions thereof may also be incorporated into compositions forcoating an implantable medical device, such as prostheses, artificialvalves, vascular grafts, stents and catheters. Accordingly, the presentinvention, in another aspect, includes a composition for coating animplantable device comprising a compound of the present invention asdescribed generally above, and in classes and subclasses herein, and acarrier suitable for coating said implantable device. In still anotheraspect, the present invention includes an implantable device coated witha composition comprising a compound of the present invention asdescribed generally above, and in classes and subclasses herein, and acarrier suitable for coating said implantable device. Suitable coatingsand the general preparation of coated implantable devices are describedin U.S. Pat. Nos. 6,099,562; 5,886,026; and 5,304,121. The coatings aretypically biocompatible polymeric materials such as a hydrogel polymer,polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylacticacid, ethylene vinyl acetate, and mixtures thereof. The coatings mayoptionally be further covered by a suitable topcoat of fluorosilicone,polysaccharides, polyethylene glycol, phospholipids or combinationsthereof to impart controlled release characteristics in the composition.

Another aspect of the invention relates to modulating CFTR activity in abiological sample or a patient (e.g., in vitro or in vivo), which methodcomprises administering to the patient, or contacting said biologicalsample with a compound of formula I or a composition comprising saidcompound. The term “biological sample”, as used herein, includes,without limitation, cell cultures or extracts thereof; biopsied materialobtained from a mammal or extracts thereof; and blood, saliva, urine,feces, semen, tears, or other body fluids or extracts thereof.

Modulation of CFTR activity in a biological sample is useful for avariety of purposes that are known to one of skill in the art. Examplesof such purposes include, but are not limited to, the study of CFTR inbiological and pathological phenomena; and the comparative evaluation ofnew modulators of CFTR.

In yet another embodiment, a method of modulating activity of an anionchannel in vitro or in vivo, is provided comprising the step ofcontacting said channel with a compound of formula (I). In preferredembodiments, the anion channel is a chloride channel or a bicarbonatechannel. In other preferred embodiments, the anion channel is a chloridechannel.

According to an alternative embodiment, the present invention provides amethod of increasing the number of functional CFTR in a membrane of acell, comprising the step of contacting said cell with a compound offormula (I). The term “functional ABC transporter” as used herein meansan CFTR that is capable of transport activity.

According to another preferred embodiment, the activity of the CFTR ismeasured by measuring the transmembrane voltage potential. Means formeasuring the voltage potential across a membrane in the biologicalsample may employ any of the known methods in the art, such as opticalmembrane potential assay or other electrophysiological methods.

The optical membrane potential assay utilizes voltage-sensitive FRETsensors described by Gonzales and Tsien (See, Gonzalez, J. E. and R. Y.Tsien (1995) “Voltage sensing by fluorescence resonance energy transferin single cells” Biophys J 69(4): 1272-80, and Gonzalez, J. E. and R. Y.Tsien (1997) “Improved indicators of cell membrane potential that usefluorescence resonance energy transfer” Chem Biol 4(4): 269-77) incombination with instrumentation for measuring fluorescence changes suchas the Voltage/Ion Probe Reader (VIPR) (see, Gonzalez, J. E., K. Oades,et al. (1999) “Cell-based assays and instrumentation for screeningion-channel targets” Drug Discov Today 4(9): 431-439).

These voltage sensitive assays are based on the change in fluorescenceresonant energy transfer (FRET) between the membrane-soluble,voltage-sensitive dye, DiSBAC₂(3), and a fluorescent phospholipid,CC2-DMFB, which is attached to the outer leaflet of the plasma membraneand acts as a FRET donor. Changes in membrane potential (V_(m)) causethe negatively charged DiSBAC₂(3) to redistribute across the plasmamembrane and the amount of energy transfer from CC2-DMPE changesaccordingly. The changes in fluorescence emission can be monitored usingVIPR™ II, which is an integrated liquid handier and fluorescent detectordesigned to conduct cell-based screens in 96- or 384-well microtiterplates.

In another aspect the present invention provides a kit for use inmeasuring the activity of a CFTR or a fragment thereof in a biologicalsample in vitro or in vivo comprising (i) a composition comprising acompound of formula (I) or any of the above embodiments; and (ii)instructions for a) contacting the composition with the biologicalsample and b) measuring activity of said CFTR or a fragment thereof. Inone embodiment, the kit further comprises instructions for a) contactingan additional composition with the biological sample; b) measuring theactivity of said CFTR or a fragment thereof in the presence of saidadditional compound, and c) comparing the activity of the CFTR in thepresence of the additional compound with the density of the CFTR in thepresence of a composition of formula (I). In preferred embodiments, thekit is used to measure the density of CFTR.

In order that the invention described herein may be more fullyunderstood, the following examples are-set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

EXAMPLES Example 11-(Benzo[d][1,3]dioxol-6-yl)-N-(5-((R)-(2-chlorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide(2) hydrochloride and1-(benzo[d][1,3]dioxol-6-yl)-N-(5-((S)-(2-chlorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide(1) hydrochloride

1-Benzo[1,3]dioxol-5-yl-cyclopropanecarboxylic acid

A mixture of benzo[1,3]dioxole-5-acetonitrile (5.10 g 31.7 mmol),1-bromo-2-chloro-ethane (9.000 mL 108.6 mmol), andbenzyltriethylammonium chloride (BTEAC, 0.181 g 0.795 mmol) was heatedto 70° C. and then 50% (wt./wt) aqueous sodium hydroxide (26 mL) wasslowly added. The reaction was stirred at 70° C. for 24 hours and thenheated to reflux (130° C. bath temperature) for 72 hours. The darkbrown/black reaction mixture was diluted with water (400 mL) andextracted twice with equal volumes ethyl acetate and dichloromethane.The basic aqueous solution was acidified with concentrated hydrochloricacid to pH less than one and the precipitate was filtered and washedwith 1 M hydrochloric acid. The solid material was dissolved indichloromethane (400 mL) and extracted twice with equal volumes of 1 Mhydrochloric acid and once with a saturated aqueous solution of sodiumchloride. The organic solution was dried over sodium sulfate andevaporated to dryness to give a white to slightly off-white solid (5.23g, 25.4 mmol, 80.1%). ESI-MS m/z calc. 206.1, found 207.1 (M+1)³⁰ .Retention time of 2.37 minutes; ¹H NMR (400 MHz, DMSO-d₆) δ 1.07-1.11(m, 2H), 1.38-1.42 (m, 2H), 5.98 (s, 2H), 6.79 (m, 2H), 6.88 (m, 1H),12.26 (s, 1H).

2-Bromo-1-(chloro-phenyl)-ethanone

Bromine (3.8 mL, 65 mmol) was added dropwise to a solution, of1-(2-chloro-phenyl)-ethanone (10 g, 65 mmol) in acetic acid (75 mL) at0° C. The mixture was then warmed to room temperature and stirredovernight. The mixture was evaporated to dryness and used in the nextstep without further purification.

N′-[5-(2-Chloro-benzoyl)-thiazol-2-yl]-N,N-dimethyl-formamidine

A mixture of thiourea (4.95 g, 65.0 mmol) anddimethoxymethyl-dimethyl-amine (23.2 g, 195 mmol) in methanol (80 mL)was heated to reflux for 30 minutes. After allowing the mixture to cool,triethylamine (19.8 g, 195 mmol) and a solution of2-bromo-1-(chloro-phenyl)-ethanone (crude from last step) in methanol(50 mL) were added. The mixture was heated to reflux for 4 hours. Thesolvent was removed and the residue was used directly in the nextprocedure.

(2-Amino-thiazol-5-yl)-(2-chloro-phenyl)-methanone

The crudeN′-[5-(2-chloro-benzoyl)-thiazol-2-yl]-N,N-dimethyl-formamidine wasdissolved in 10% HCl (150 mL) and heated to 70° C. for 4 hours. Theprecipitate was filtered, washed with ether, and then suspended in a 10%sodium carbonate solution (250 mL). The suspension was stirred for 1hour and the precipitate was filtered, washed with ether, and dried inair to give (2-amino-thiazol-5-yl)-(2-chloro-phenyl)-methanone as abrown solid (8.5 g, 36 mmol, 55% from 1-(2-chloro-phenyl)-ethanone). ¹HNMR (400 MHz, DMSO-d₆) δ 7.27 (s, 1H), 7.41-7.58 (m, 4H), 8.37 (s, 2H).ESI-MS m/z calc. 238.0, found; 239.3 (M+1)³⁰.

1-Benzo[1,3]dioxol-5-yl-cyclopropanecarboxylic acid[5-(2-chloro-benzoyl)-thiazol-2-yl]amide

1-Benzo[1,3]dioxol-5-yl-cyclopropanecarboxylic acid (1.29 g, 6.28 mmol)was placed in an even-dried flask under nitrogen. Thionyl chloride (3mL) and N,N-dimethylformamide (0.3 mL) were added and the solution wasallowed to stir for 2 hours. The excess thionyl chloride was removedunder vacuum and the resulting solid was suspended in 30 ml of anhydrous1,4-dioxane containing triethylamine (1.77 mL, 12.6 mmol).(2-Amino-thiazol-5-yl)-(2-chloro-phenyl)-methadone (1.50 g, 6.28 mmol)suspended in 10 ml of anhydrous 1,4-dioxane was slowly added to thissuspension. The resulting suspension was allowed to stir for 20 minutes.The mixture was filtered and then the filtrate was evaporated todryness. The crude product was dissolved in 50 mL of dichloromethane andwashed three times with 50 mL of 1N HCl, saturated aqueous sodiumbicarbonate, and saturated aqueous sodium chloride. The organic layerwas then dried over sodium sulfate and evaporated to dryness to yieldthe product as a beige solid (1.51 g, 3.54 mmol, 56.4%). ¹H NMR (400MHz, DMSO-d₆) δ 1.20-1.24 (m, 2H), 1.54-1.57 (m, 2H), 6.01 (s, 2H), 6.88(d, J=1.3 Hz, 2H), 6.98 (s, 1H), 7.48-7.52 (m, 1H), 7.56-7.60 (m, 3H),7.77 (s, 1H), 11.98 (s, 1H). ESI-MS m/z calc. 426.0, found; 427.3(M+1)⁺; Retention time 3.46 minutes.

1-Benzo[d][1,3]dioxol-6-yl)-N-(5-((2-chlorophenyl)(hydroxy)methyl)thiazol-2-yl)-cyclopropanecarboxamide

1-Benzo[1,3]dioxol-5-yl-cyclopropanecarboxylic acid[5-(2-chloro-benzoyl)-thiazol-2-yl]-amide (1.0 g, 2.3 mmol) wassuspended in 150 mL of anhydrous methanol. Sodium borohydride (1.3 g, 35mmol) was slowly added and the resulting pale yellow solution wasallowed to stir for 1 hour at room temperature. The crude product wasevaporated to dryness and then dissolved in a minimum of ethyl acetate.The organic was washed three times with an equal volume of 1Nhydrochloric acid, saturated aqueous sodium bicarbonate, and saturatedaqueous sodium chloride. The organic layer was then dried over sodiumsulfate, filtered, and evaporated to dryness to yield the product as abeige solid (0.64 g, 1.5 mmol, 63%). ¹H NMR (400 MHz, DMSO-d₆) δ1.10-1.14 (m, 2H), 1.41-1.45 (m, 2H), 6.00 (s, 2H), 6.14 (s, 1H), 6.86(d, J=1.0 Hz, 2H), 6.95 (t, J=1.0 Hz, 1H), 7.14 (d, J=0.6 Hz, 1H),7.29-7.34 (m, 1H), 7.38-7.43 (m, 2H), 7.71 (d, J=7.5 Hz, 1H), 10.93 (s,1H) ESI-MS m/z calc. 428.1, found; 429.5 (M+1)⁺ Retention time 3.17minutes.

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamide

1-Benzo[1,3]dioxol-5-yl-cyclopropanecarboxylic acid[5-[(2-chloro-phenyl)-hydroxy-methyl]-thiazol-2-yl]-amide (0.500 g, 1.17mmol) was placed in 10 mL of anhydrous dichloromethane containingtrieihylamine (984 μL, 7.02 mmol). The mixture was cooled to 0° C. andmethanesulfonyl chloride (364 μL, 4.68 mmol) was added, immediatelyfollowed by (R)-pyrrolidin-3-ol (945 μL, 11.7 mmol) and the solution wasallowed to stir for 10 minutes at room temperature. The crude productwas washed three times with an equal volume of a saturated aqueoussolution of sodium bicarbonate, followed by a saturated aqueous solutionof sodium chloride. The organic layer was then dried over sodium sulfateand evaporated to dryness. The crude mixture was purified by column,chromatography (20-90% ethyl acetate in hexanes on silica gel) to yieldthe product as a white solid (194.2 mg, 0.390 mol, 33.3%). ¹H NMR (400MHz, DMSO-d₆) δ 1.09-1.15 (m, 2H), 1.41-1.44 (m, 2H), 1.54-1.63 (m, 1H),1.93-2.77 (m, 5H), 4.20 (S, 1H), 4.72-4.77 (m, 1H), 4.96 (d, J=7.0 Hz,1H), 6.00 (s, 2H), 6.85 (d, J=0.9 Hz, 2H), 6.95 (s, 1H), 7.24-7.29 (m,1H), 7.37-7.43 (m, 3H), 7.76-7.80 (m, 1H), 11.03 (s, 1H. ESI-MS m/zcalc. 497.1, found; 498.1 (M+1)⁺; Retention time 2.36 minutes.

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamidehydrochloride

A solution of HCl in ether (0.1556 mL, 0.3112 mmol, 1M) was slowly addedto a stirred solution of1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamide(0.1550 g, 0.311.2 mmol) in 100 mL of anhydrous dichloromethane. Thesolution was evaporated to dryness to give the pure product (0.1654 g,0.3095, 99.45%). ESI-MS m/z calc, 497.1, found; 498.1 (M+1)⁺; Retentiontime 5.74 minutes.

1-(Benzo[d][1,3]dioxol-6-yl)-N-(5-((S)-(2-chlorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamide(1) hydrochloride and1-(Benzo[d][1,3]dioxol-6-yl)-N-(5-((R)-(2-chlorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamide(2) hydrochloride

1-(Benzo[d][1,3]dioxol-6-yl)-N-(5-((2-chlorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamidehydrochloride prepared above was separated using utilizing a ChiralpakAS-H 4.6 mm×250 mm column from Chiral Technologies.

20-25 μL of a 2 mg/mL solution of1-(benzo[d][1,3]dioxol-6-yl)-N-(5-((2-chlorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamidehydrochloride in methanol was injected onto the Chiralpak AS-H columnand was eluted with a mixture of 10% of a 50/50 (v/v) mix-lure ofethanol and methanol in hexanes at 1.5 mL/min.

The first eluting product under these conditions had a retention time of8.2 min (Chiralpak AS-H column). This product had a retention time 14.5min. on a Chiralpak OJ-H 4.6 mm×250 mm column (25% of a 50/50 (v/v)mixture of ethanol and methanol in hexanes at 1.0 mL/min.)

The second product elated at 9.6 min using a Chiralpak AS-H column. Thissecond product had a retention time of 10.9 mm. on a Chiralpak OJ-H 4.6mm×250 mm column (25% of a 50/50 (v/v) mixture of ethanol and methanol mhexanes at 1.0 mL/min.)

Example 2(R)-1-((2-(1-(Benzo[d][1,3]dioxol-6-yl)cyclopropanecarboxamido)thiazol-5-yl)(2-chlorophenyl)methyl)pyrrolidin-3-yl(1S,2R,5S)-2-isopropyl-5-methylcyclohexyl carbonate

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamide(3.00 g, 6.02 mmol) was suspended in 200 mL of anhydrous dichloromethanecontaining N,N-dimethylpyridin-4-amine (2.20 g, 18.0 mmol),(1S,2R,5S)-2-Isopropyl-5-methylcyclohexyl chloroformate (1.91 mL, 9.00mmol) was slowly added to the suspension and the resulting mixture washeated to reflux for 16 hours. The resulting pale yellow solution wasallowed to cool to room temperature, diluted with 20 mL of methanol, andthen evaporated to dryness. The crude reaction mixture was separated on330 g of silica gel utilizing a gradient of 0-5% methanol indicholoromethane to yield the pure product as a pale yellow solid (2.008g, 2.9529 mmol, 49.1%). ESI-MS m/z calc. 679.3, found; 680.5 (M+1)⁺;Retention time 3.88 minutes.

(R)-1-((S)-(2-(1-(Benzo[d][1,3]dioxol-6-yl)cyclopropanecarboxamido)thiazol-5-yl)(2-chlorophenyl)methyl)pyrrolidin-3-yl(1S,2R,5S)-2-isopropyl-5-methylcyclohexyl carbonate and(R)-1-((R)-(2-(1-(benzo[d][1,3]dioxol-6-yl)cyclopropanecarboxamido)thiazol-5-yl)(2-chlorophenyl)methyl)pyrrolidin-3-yl(1S,2R,5S)-2-isopropyl-5-methylcyclohexyl carbonate

(R)-1-(2-(1-(Benzo[d][1,3]dioxol-6-yl)cyclopropanecarboxamido)thiazol-5-yl)(2-chlorophenyl)methyl)pyrrolidin-3-yl(1S,2R,5S)-2-isopropyl-5-methylcyclohexyl carbonate prepared above wasseparated utilizing a Chiralpak AD 21 mm×250 mm column from ChiralTechnologies. 1 mL of a 30 mg/mL solution of compound prepared above inisopropanol was injected onto the Chiralpak AD column and was elutedwith a mixture of 7.5% isopropanol in heptane at 15 mL/min. The firsteluting product(R)-1-((S)-(2-(1-(benzo[d][1,3]dioxol-6-yl)cyclopropanecarboxamido)thiazol-5-yl)(2-chlorophenyl)methyl)pyrrolidin-3-yl(1S,2R,5S)-2-isopropyl-5-methylcyclohexyl carbonate) had a retentiontime of 14.4 Min. (Chiralpak AD column); ¹H NMR (400 MHz, CD₃OD) δ 0.79(d, J=7.0 Hz, 3H), 0.85-1.84 (m, 18H), 1.86-2.04 (m, 3H), 2.23-2.34 (m,1H), 2.43-2.51 (m, 1H), 2.62-2.69 (m, 1H), 2.73-2.78 (m, 2H), 4.45-4.54(m, 1H), 5.04-5.08 (m, 1H), 5.12 (s, 1H), 5.99 (s, 2H), 6.82 (d, J=8.5Hz, 1H), 6.93-6.97 (m, 2H), 7.25 (t, J=6.8 Hz, 1H), 7.32 (s, 1H),7.34-7.40 (m, 2H), 7.85 (dd, J=8.1, 1.6 Hz, 1H). ESI-MS m/z calc. 679.3,found: 680.5 (M+1)⁺; Retention time 3.91 minutes.

The second eluting product(R)-1-((R)-(2-(1-(benzo[d][1,3]dioxol-6-yl)cyclopropanecarboxamido)thiazol-5-yl)(2-chlorophenyl)methyl)pyrrolidin-3-yl(1S,2R,5S)-2-isopropyl-5-methylcyclohexyl carbonate) had a retentiontime of 28.6 mm (Chiralpak AD column). ESI-MS m/z calc. 679.3, found;680.5 (M+1)⁺; Retention time 3.86 minutes.

1-(Benzo[d][1,3]dioxol-6-yl)-N-(5-((S)-(2-chlorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamidehydrochloride and1-(Benzo[d][1,3]dioxol-6-yl)-N-(5-((R)-(2-chlorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamidehydrochloride

The first eluting product from previous step (1.20 g, 1.76 mmol) wasstirred at room temperature in 182 mL of methanol containing (0.9153 g,16.31 mmol) of potassium hydroxide for 4 days. The solution was thencooled to 0° C. and 16.31 mL of 1N HCl was slowly added to the reactionmixture. The resulting solution was evaporated to near dryness and thenpartitioned between 100 mL of dichloromethane and 100 mL of a saturatedaqueous solution of sodium bicarbonate. The layers were separated andthe organic layer was washed twice with and equal volume of a saturatedaqueous solution of sodium bicarbonate followed by three washes with asaturated aqueous solution of sodium chloride. The organic layer wasthen dried over anhydrous sodium sulfate, concentrated, and purified on120 g of silica gel utilizing a gradient of 20-100% ethyl acetate inhexanes over 40 minutes to yield the pure product as a white solid (0.70g, 1.4 mmol). This material was then dissolved in a minimum ofdichloromethane and 1.4 mL of 1M HCl in ether was added to the solution.The solution was evaporated to dryness to yield the HCl salt as a whitesolid (0.7818 g, 1.463 mmol, 83.1%). ESI-MS m/z calc, 497.1, found;498.3 (M+1)⁺; Retention time 2.36 minutes. Retention time of thisproduct was 14.5 min. on a Chiralpak OJ-H 4.6 mm×250 mm column (25% of a50/50 (v/v) mixture of ethanol and methanol in hexanes at 1.0 mL/min).

¹H NMR (400 MHz, DMSO-d₆) δ (free base ) 1.07-1.15 (m, 2H), 1.40-1.44(m, 2H), 1.51-1.65 (m, 1H), 1.95-2.08 (m, 1H), 2.32-2.58 (m, 4H),4.10-4.31 (m, 1H), 4.76 (d, J=4.3 Hz, 1H), 4.95 (s, 1H), 6.00 (s, 2H),6.85 (s, 2H), 6.95 (d, J=0.9 Hz, 1H), 7.26 (t, J=7.6 Hz, 1H), 7.34-7.44(m, 3H), 7.78 (d, J=7.7 Hz, 1H), 11.02 (s, 1H)

The second during product torn previous step (0.1018 g, 0,1497 mmol) wasstirred at room temperature in 15 mL of methanol containing (9.5 mg,0.17 mmol) of potassium hydroxide for 3 days. An additional, aliquot ofpotassium hydroxide was added (64.8 mg, 1.15 mmol) and the solution wasallowed to stir for and additional 3 days. The solution was then cooledto 0° C. and 1.319 mL of 1N HCl was slowly added to the reactionmixture. The resulting solution was evaporated to near dryness and thenpartitioned between 10 mL of dichloromethane and 10 mL of a saturatedaqueous solution of sodium chloride. The organic layer was then driedover anhydrous sodium sulfate, concentrated, and purified on 12 g ofsilica gel utilizing a gradient of 20-100% ethyl acetate in hexanes over40 minutes. The column was then flushed with ethylacetate containing2.5% trietlrylamine. The pure fractions were combined and evaporated todryness to yield the pure product as a white solid (15.7 mg, 0.0315mmol). This material was then dissolved in a minimum of dichloromethaneand 0.0315 mL of 1M HCl in ether was added to the solution. The solutionwas evaporated to dryness to yield the HCl salt as a white solid (16.8g, 0.0315 mmol, 21.0%), ESI-MS m/z calc. 497.1, found; 498.3 (M+1)⁺;Retention time 2.42 minutes. Retention time 10.9 minutes on a ChiralpakOJ-H 4.6 mm×250 mm column (25% of a 50/50 (v/v) mixture of ethanol andmethanol in hexanes at 1.0 mL/min.)

(R)-N-(2-Chlorobenzylidene)-1,1-dimethylethylsulfinamide

To a stirred anhydrous dioxane (500 mL) was added 2-chlorobenzaldehyde(34.8 g, 247.5 mmol) and the solution was cooled down to 0° C. in an icebath. A solution of (R)-2-methylpropanesulfinamide (30.0 g, 247.5 mmol)in anhydrous dioxane (100 mL) was added to the aldehyde solution.Ti(OPr)₄ (105.5 g, 371.3 mmol) was then slowly added to the solutionwhile stirring at 0° C. The reaction mixture was allowed to warm up to25° C., stirred at 25° C. for 18 h, quenched with NaHCO₃ and thenfiltered through a short plug of Celite using EtOAc. The organic layerwas separated from the aqueous layer and dried over Na₂SO₄ andconcentrated. The crude product was purified by column chromatography(0-25% EtOAc/Hexane) to provide(R)-N-(2-chlorobenzylidene)-1,1-dimethylethylsulfinamide as a yellowliquid (45.1 g, 75%). ¹H NMR (400 MHz, CDCl3) δ 9.05 (s, 1H), 8.06 (dd,J=7.9, 1.1 Hz, 1H), 7.47-7.41 (m, 2H), 7.37-7.33 (m, 1H), 1.28 (s, 9H).HPLC ret. time 3.45 min, 10-99% CH₃CN, 5 min run; ESI-MS 244,3 m/z(MH⁺).

tert-Butyl thiazol-2-ylcarbamate

To a solution of 2-aminothiazole (20.0 g, 199.7 mmol) and (Boc)₂O (48.0g, 219.7 mmol) in anhydrous THF (100 mL) were added DMAP (20 mg) andEt₃N (36.0 mL, 260.0 mmol). The reaction mixture was stirred at 25° C.for 18 h, diluted with CH₂Cl₂ and washed with 0.1 N HCl (X 1), brine(X 1) and H₂O (X 1). The organic layer was separated from the aqueouslayer, dried over Na₂SO₄ and concentrated. The crude product waspurified by column chromatography (0-40% EtOAc/Hexane) to providetert-butyl thiazol-2-ylcarbamate as a white solid (20.7 g, 72%). ¹H-NMR(400MHz, CDCl₃) δ 11.44 (s, 1H), 738 (d, J=3.6 Hz, 1H), 6.89 (d, J=3.6Hz, 1H), 1.58 (s, 9H). HPLC ret. time 2.61 min, 10-99% CH₃CN, 5 min run;ESI-MS 145.1 m/z (MH⁺).

(R)-N-(1-((2-t-butoxylcarbonylamino)thiazol-5-yl)-1-(2-chlorophenyl)-methyl)-1,1-dimethylethylsulfinamide

A solution of tert-butyl thiazol-2-ylcarbamate (15.0 g, 75.0 mmol) inanhydrous THF (175 mL) was stirred and cooled down to −78° C. To thissolution was slowly added n-BuLi (2.5 M in hexane: 60.0 mL, 150.0 mmol).Upon completion of m-BuLi addition, the mixture was allowed to warm upto −40° C., maintained at −40° C. for 1 h and then cooled down to −78°C. A solution of(R)-N-(2-chlorobenzylidene)-1,1-dimethylethylsulfinamide (10.0 g, 41.0mmol) in anhydrous THF (175 mL) previously cooled to −78° C. was slowlyadded to the above solution via canulation. The reaction was kept at−78° C. for 0.5 h, allowed to warm up to room temperature and stirred atroom temperature for 2 h. The reaction was then quenched with aqueousNH₄Cl, and the crude product was extracted with EtOAC (X 3). Thecombined organic layers were dried over Na₂SO₄ and concentrated. Thecrude product was purified by column chromatography (0-80% EtOAc/Hexane)to provide a diastereomeric mixture of(R)-N-(1-((2-t-butoxylcarbonylamino)thiazol-5-yl)-1-(2-chlorophenyl)-methyl)-1,1-dimethylethylsulfinamideas a yellow solid (15.1 g, 83%) that was used directly in the next step.

(R)-N-(S)-(1-((2-Aminothiazol-5-yl)-1-(2-chlorophenyl)methyl)-1,1-dimethylethylsulfinamideand(R)-N-(R)-(1-((2-aminothiazol-5-yl)-1-(2-chlorophenyl)methyl)-1,1-dimethylethylsulfinamide

To a solution of of(R)-N-(1-((2-t-butoxylcarbonylamino)thiazol-5-yl)-1-(2-chlorophenyl)-methyl)-1,1-dimethylethylsulfinamide(7.0 g, 15.8 mmol) in CH₂Cl₂ (28 mL) was added trifluoroacetic acid (28mL). The reaction was stirred at room temperature for 3.5 h. Thetrifluoroacetic acid and CH₂Cl₂ were removed under vacuum. The crudeproduct was re-dissolved in CH₂Cl₂, washed with aqueous NaHCO₃ (20 ml×2)and water (20 mL×1), dried over Na₂So₄ and concentrated. The crudeproduct was purified by column chromatography (0-5% EtOH/EtOAc) toprovide(R)-N-(S)-1-(2-aminothiazol-5-yl)-1-(2-chlorophenyl)methyl)-1,1-dimethylethylsulfinamideand(R)-N-(R)-1-(2-aminothiazol-5-yl)-1-(2-chlorophenyl)methyl)-1,1-dimethylethylsulfinamide.

(R)-N-(S)-1-(2-aminothiazol-5-yl)-1-(2-chlorophenyl)methyl)-1,1-dimethylethylsulfinamide:yellow solid, 4.3 g (79%), ¹H-NMR (400 MHz, CDCl₃) δ 7.53 (dd, J=7.6,1.8 Hz, 1H), 7.38 (dd, J=7.7, 1.5 Hz, 1H), 7.32 (td, J=7.5, 1.6 Hz, 1H),7.27 (td, J=7.5, 1.8 Hz, 1H), 6.84 (d, J=0.7 Hz, 1H), 6.17 (s, 2H), 6.07(d, J=4.5 Hz, 1H), 4.17 (d, J=4.6 Hz, 1H), 1.26 (s, 9H). HPLC ret. time2.11 min, 10-99% CH₃CN, 5 min run; ESI-MS 344.0 m/z (MH⁺).

(R)-N-(R)-1-(2-Aminothiazol-5-yl)-1-(2-chlorophenyl)methyl)-1,1-dimethylethylsulfinamide:yellow solid, 596 mg, (11%). HPLC ret. time 2.35 min, 10-99% CH₃CN, 5min run; ESI-MS 344.0 m/z (MH⁺).

(S)-N-(5-((R)-t-butylsulfinylamino(2-chlorophenyl)methyl)thiazol-2yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

To 1-(benzo[d][1,3]dioxol-5-yl-cyclopropanecarboxylic acid (2.16 g, 10.5mmol) was slowly added SOCl₂(2.3 mL, 31.5 mmol) followed by DMF (3drops). The reaction mixture was heated at 60° C. for 0.5 h. The excessSOCl₂was removed under vacuum. The acid chloride (10.5 mmol) was thendissolved in anhydrous CH₂Cl₂(16 mL) and was slowly added to a cold(temperature ˜0° C.) solution of(R)-N-(S)-1-(2-aminothiazol-5-yl)-1-(2-chlorophenyl)methyl)-1,1-dimethylethylsulfinamide(3.6 g, 10.5 mmol) and Et₃N (7.33 mL, 52.6 mmol) in anhydrous CH₂C₂ (16mL). The reaction mixture was stirred at 25° C. for 18 h, diluted withCH₂Cl₂ and washed with 1 N HCl (50 mL×2), NaHCO₃ (50 mL×1) and brine (50mL×1). The organic layer was separated from the aqueous layer, driedover Na₂SO₄ and concentrated. The crude product was purified by columnchromatography (0-80% EtOAc/Hexane) to provide(S)-N-(5-((R)-t-butylsulfinylamino-(2-chlorophenyl)-methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideas a yellow solid (4.7 g, 84%), ¹H-NMR (400 MHz, CDCl₃) δ 8.52 (s, 1H),7.60 (dd, J=7.7, 1.7 Hz, 1H), 7.35 (dd, J=7.8, 1.4 Hz, 1H), 7.32 (dd,J=7.5, 1.4 Hz, 1H), 7.29 (d, J=0.7 Hz, 1H), 7.23 (dd, J=7.6, 1.7 Hz,1H), 6.89 (dd, J=7.9, 1.8 Hz, 1H), 6.86 (d, J=1.6 Hz, 1H), 6.81 (d,J=7.9 Hz, 6.21 (d, J=4.0 Hz, 1H), 6.01 (s, 2H), 3.93 (d, J=4.0 Hz, 1H),1.75-1.66 (m, 2H), 1.27 (s, 9H), 1.22 (t, J=3.3 Hz, 2H), HPLC ret. time3.52 min, 10-99% CH₃CN, 5 min run; ESI-MS 532.0 m/z (MH⁺).

(S)-N-(5-(Amino-(2-chlorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

To a solution of(S)-N-(5-((R)-t-butylsulfinylamino-(2-chlorophenyl)-methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropane-carboxamide(11.25 g, 21.19 mmol) in MeOH (100 mL) was added 4 M HCl in dioxane (32mL, 128 mmol). The reaction mixture was stirred at 25° C. for 1.5 h andevaporated to dryness. The crude product was dissolved in CH₂Cl₂. Theorganic layer was washed with aqueous NaHCO₃(50 mL×2), brine (50 mL×1),dried over Na₂So₄ and concentrated. The crude product was purified bycolumn chromatography (0-2.5% Et₃N-EtOAc) to provide(S)-N-(5-(amino(2-chlorophenyl)-methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(6.5 g, 72%, >99% ee). ¹H-NMR (400 MHz, CDCl₃) δ 8.51 (s, 1H), 7.60 (dd,J=7.7, 1.7 Hz, 1H), 7.33 (dd, J=7.9, 1.3 Hz, 1H), 7.29-7.25 (m, 1H),7.20 (td, J=7.6, 1.7 Hz, 1H), 7.16 (d, J=1.0 Hz, 1H), 6.89 (td, J=7.8,1.7 Hz, 2H), 6.80 (d, J=7.9 Hz, 1H), 6.01 (s, 2H), 5.79 (s, 1H), 1.86(bs, 2H), 1.72-1.69 (m, 2H), 1.22-1.19 (m, 2H). HPLC ret. time 2.66 min,10-99% CH₃CN, 5 min run; ESI-MS 428.1 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chlorophenyl)((R)-3-dimethyl-t-butylsilylhydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamide

To a solution of(S)-N-(5-(amino-(2-chlorophenyl)-methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(214 mg, 0.5 mmol) in MeOH (2.5 mL) was added(R)-4-chloro-3-dimethyl-t-butylhydroxybutanal (14.2 mg, 0.6 mmol). Thereaction mixture was stored at 25° C. for 5 min before NaBH₄ (28 mg,0.75 mmol) was added. Stirring was continued at 25° C. for 1 h. Thereaction was diluted with H₂O and extracted with EtOAc. The combinedorganic layers was washed with brine and dried over MgSO4. After theremoval of solvent, the residue was purified by column chromatography(10-20% EtOAc-Hexane) to afford1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chlorophenyl)((R)-3-dimethyl-t-butylsilylhydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamide(162 mg, 53%).

¹H-NMR (400 MHz, CDCl₃) δ 8.45 (s, 1H), 7.81 (dd, J=8.1, 1.6 Hz, 1H),7.28 (s, 1H), 7,25-7.22 (m, 2H), 7.13-7.09 (m, 1H), 6.85 (id, J=7.7, 1,7Hz, 2H), 6.77 (d, J=7.9 Hz, 1H), 5,.8 (s, 2H), 5.05 (s, 1H), 4.36-4.31(m, 1H), 2.81 (dd, J=9.8, 6.2 Hz, 1H), 2.57-2.46 (m, 2H), 2.37 (dd,J=9.8, 4.5 Hz, 1H), 2.08-1.99 (m, 1H), 1.73-1.62 (m, 3H), 1.17 (t, J=3.9Hz, 2H), 0.85 (s, 9H), −0.01 (d, J=6.9 Hz, 6H), HPLC ret. time 3.51 min,10-99% CH₃CN, 5 min run; ESI-MS 612.41 m/x (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chlorophenyl)((R)-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamide(1)

A mixture of1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chlorophenyl)((R)-3-dimethyl-t-butylsilylhydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamide(61 mg. 0.1 mmol) and TBAF (1 M in THF, 0.6 mL, 0.6 mmol) was stirred at25° C. for 3 h. The reaction was diluted with H₂O and extracted withEtOAc. The combined organic layers was washed with brine and dried overMgSO₄After the removal of solvent, the residue was purified by columnchromatography (10-20% EtOAc-Hexane) to afford1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chlorophenyl)((R)-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-cyclopropanecarboxamide(1) (30 mg, 62%, >99% ee). ¹H-NMR (400 MHz, DMSO-d₆) δ 11.00 (s, 1H),7.79 (dd, J−7.8, 1.5 Hz, 1H), 7.43-7.37 (m, 3H), 7.26 (td, J=7.0, 1.4Hz, 1H), 6.95 (s, 1H), 6.86 (d, J=0.7 Hz, 2H), 6.00 (s, 2H), 4.96 (s,1H), 4.76 (d, J=4.4 Hz, 1H), 4.20 (ddd, J=6.6, 3.4 Hz, 1H), 2.56-2.45(m, 2H), 2.43-2.36 (m, 2H), 2.06-1.97 (m, 1H), 1.62-1.54 (m, 1H), 1.43(q, J=3.7 Hz, 2H), 1.14-1.09 (m, 2H). HPLC ret time 2.85 min, 10-99%CH₃CN, 5 min run; ESI-MS 498.0 m/z (MH⁺).

(R)-N-(5-((R)-t-butylsulfinylamino(2-chlorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide

(R)-N-(5-((R)-t-butylsulfinylamino(2-chlorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamidewas prepared from(R)-N-(R)-1-(2-aminothiazol-5-yl)-1-(2-chlorophenyl)methyl)-1,1-dimethylethylsulfinamideand 1-(benzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxylic acid using thesame protocol for(S)-N-(5-((R)-t-butylsulfinylamino(2-chlorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide.¹H-NMR (400 MHz, CDC₃) δ 8.55 (s, 1H), 7.62 (dd, J=7.7, 1.7 Hz, 1H),7.37 (d, J=0.7 Hz, 1H), 7.34 (dd, J=7.8, 1.4 Hz, 1H), 7.31-7.20 (m, 2H),6.87 (td, J=8.4, 1.7 Hz, 2H), 6.81 (d, J=7.9 Hz, 1H), 6.30 (d, J=2.8 Hz,1H), 6.01 (s, 2H), 3.74 (d, J=2.8 Hz, 1H), 1.71-1.68 (m, 2H), 1.24 (s,9H), 0.88 (t, J=69 Hz, 2H), HPLC ret. time 3.59 mm, 10-99% CH₃CN, 5 minrun; ESI-MS 532.1 m/z (MH⁺).

(R)-N-(5-(Amino(2-chlorophenyl)-methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

(R)-N-(5-(Amino(2-chlorophenyl)-methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamidewas prepared from(R)-N-(5-((R)-t-butylsulfinylamino(2-chlorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideusing the same protocol described for(S)-N-(5-(amino(2-chlorophenyl)-methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide.¹H-NMR (400 MHz, CDCl₃) δ 8.47 (s, 1H), 7.60 (dd, J=7.7, 1.7 Hz, 1H),7.33 (dd, J=7.9, 1.3 Hz, 1H), 7.27 (td, J=7.4, 1.5 Hz, 1H), 7.20 (td,J=7.6, 1.8 Hz, 1H), 7.16 (d, J=1.0 Hz, 1H), 6.89 (td, J=8.2, 1.7 Hz,2H), 6.81 (d, J=7.9 Hz, 1H), 6.01 (s, 2H), 5.79 (s, 1H), 1.89 (s, 2H),1.72-1.70 (m, 2H), 1,22-1.19 (m, 2H). HPLC ret. time 2.52 min, 10-99%CH₃CN, 5 min run; ESI-MS 428.2 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(2-chlorophenyl)((R)-3-dimethyl-t-butylsilylhydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(2-chlorophenyl)((R)-3-dimethyl-t-butylsilylhydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamidewas prepared from(R)-N-(5-(amino(2-chlorophenyl)-methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideusing the same protocol for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chlorophenyl)((R)-3-dimethyl-t-butylsilylhydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide.1H-NMR (400 MHz, CDCl₃) δ 8.45 (s, 1H), 7.79 (dd, J=8.1, 1.5 Hz, 1H),7.28-7.23 (m, 3H), 7.14-7.10 (m, 1H), 6.85 (td, J=8.1, 1.7 Hz, 2H), 6.78(d, J=7.9 Hz, 1H), 5.99 (s, 2H), 5.08 (s, 1H), 4.36-4.31 (m, 1H), 2.94(dd, J=9.9, 6.3 Hz, 1H), 2.65 (td, J=8.4, 3.9 Hz, 1H), 2.56 (q, J=8.3Hz, 1H), 2.16 (dd, J=9.9, 4.6 Hz, 1H), 2.06-1.97 (m, 1H), 1.73-1.62 (m,3H), 1.20-1.15 (m, 2H), 0.84 (s, 9H), −0.01 (d, J=7.9 Hz, 6H). HPLC ret.time 3.51 min, 10-99% CH₃CN, 5 min run; ESI-MS 612.41 m/z (MH+).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(2-chlorophenyl)((R)-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(2-chlorophenyl)((R)-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamidewas prepared from1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(2-chlorophenyl)((R)-3-dimethyl-t-butylsilylhydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamideusing the same protocol described for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chlorophenyl)((R)-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide.¹H-NMR (400 MHz, DMSO-d6) δ 11.01 (s, 1H), 7.79 (dd, J=7.8, 1.6 Hz, 1H),7.45-7.39 (m, 3H), 7.30-7.26 (m, 1H), 6.97 (s, 1H), 6.87 (d, J=0.9 Hz,2H), 6.02 (s, 2H), 4.99 (s, 1H), 4.73 (d, J=4.3 Hz, 1H), 4.24-4.18 (m,1H), 2.72 (dd, J=9.9, 6.1 Hz, 1H), 2.62 (q, J=7.8 Hz, 1H), 2.37-2.32 (m,1H), 2.22 (dd, J=9.9, 3.2 Hz, 1H), 2.04-1.96 (m, 1H), 1.64-1.57 (m, 1H),1.44 (q, J=3.8 Hz, 2H), 1.13 (t, J=3.8 Hz, 2H). HPLC ret. time 2.56 min,10-99% CH₃CN, 5 min run; ESI-MS 498.3 m/z (MH⁺).

The methods outlined in Scheme A, Scheme B, and Scheme C below were usedto make representative compounds of this invention as recited below.

1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid

A mixture of 2-(benzo[d][1,3]dioxol-5-yl)acetonitrile (5.10 g, 31.7mmol), 1-bromo-2-chloro-ethane (9.000 mL 108.6 mmol), andbenzytriethylammonium chloride (BTEAC, 0.181 g, 0.795 mmol) was heatedto 70° C. and then 50% (wt./wt.) aqueous sodium hydroxide (26 ml) wasslowly added. The reaction was stirred at 70° C. for 88 hours and thenheated, to reflux (130° C. bath temperature) for 24 hours. The darkbrown/black reaction mixture was diluted with water (400 mL) andextracted twice with equal volumes of ethyl acetate and dichloromethane.The basic aqueous solution was acidified with concentrated hydrochloricacid to pH less than one and the precipitate was filtered and washedwith 1 M hydrochloric acid. The solid material was dissolved indichloromethane (400 mL) and washed twice with equal volumes of 1 Mhydrochloric acid and once with brine. The organic solution was driedover sodium sulfate and evaporated to dryness to give a white toslightly off-white solid (5.23 g, 80.1%). ¹H NMR (400 MHz, DMSO-d₆) δ1.07-1.11 (m, 2H), 1.38-1.42 (m, 2H), 5.98 (s, 2H), 6.79 (m, 2H), 6.88(m, 1H), 12.26 (s, 1H); HPLC ret. time 2.37 min, 10-99% CH₃CN, 5 minrun; ESI-MS 206.1 m/z (MH⁺).

Methyl 2,2-difluorobenzo[d][1,3]dioxole-5-carboxylate

A solution of 5-bromo-2,2-difluorobenzo[d][1,3]dioxole (11.8 g, 50.0mmol) and tetrakis(triphenylphosphine)palladium (0) [5.78 g, 5.00 mmol]in methanol (20 mL) containing acetonitrile (30 mL) and triethylamine(10 mL) was stirred under a carbon monoxide atmosphere (55 psi) at 75°C. (oil bath temperature) for 15 h. The cooled reaction mixture wasfiltered and the filtrate was evaporated to dryness. The residue waspurified by silica gel column chromatography to give methyl2,2-difluorobenzo[d][1,3]dioxole-5-carboxylate (11.5 g), which was useddirectly in the next step.

(2,2-Difluorobenzo[d][1,3]dioxol-5-yl)methanol

Method A: Methyl 2,2-difluorobenzo[d][1,3]dioxol-5carboxylate fromprevious step (11.5 g) dissolved in anhydrous tetrahydrofuran (20 mL)was slowly added to a suspension of lithium aluminum hydride (4.10 g,106 mmol.) in anhydrous THF (100 mL) at 0° C. The mixture was thenwarmed to room temperature and stirred at for 1 h. The reaction mixturewas cooled to 0° C. and treated with water (4.1 g), followed by sodiumhydroxide (10% aqueous solution, 4.1 mL). The resulting slurry wasfiltered and washed with THF. The combined filtrate was evaporated todryness and the residue was purified by silica gel column chromatographyto give (2,2-difluorobenzo[d][1,3]dioxol-5-yl)methanol as a colorlessoil (7.2 g, 76% over two steps).

Method B: To a solution of 2,2-difluorobenzo[d][1,3]dioxol-5carbaldehyde(125 g, 0.67 mol) in anhydrous THF (400 mL) was added NaBh₄ (28 g, 0.74mol) in portions at 0° C. The mixture was stirred for 1 h at 0° C., thenpoured into 500 mL of water. The mixture was extracted with ethylacetate (200 mL×3). The organic phase was dried over Na₂So₄ andconcentrated in vacuo to give(2,2-difluorobenzo[d][1,3]dioxol-5-yl)methanol as colorless oil (120 g,95%).

Method C: To a suspension of LAH (12.6 g, 0.33 mol) in THF (100 mL) wasadded dropwise a solution of 2,2-difluorobenzo-1,3dioxole-5-carboxylicacid (30 g, 0.15 mol) in THF (200 mL) at 0□ under N₂. The mixture wasallowed to warm to room temperature and stirred for one hour at thistemperature. Then, water (12.6 g) and aq. NaOH (10%, 12.6 g) were addeddropwise at 0□. The resulting mixture was filtered. The filtrate wasdried over anhydrous Na₂SO₄ and evaporated under vacuum to give crude(2,2-difluoro-1,3-benzodioxol-5-yl)methanol (25.5 g, 91.3%), which wasused directly in the next step.

5-(Chloromethyl)-2,2-difluorobenzo[d][1,3]dioxol

A solution of (2,2-difluorobenzo[d][1,3]dioxol-5-yl)methanol (120 g,0.64 mol) in neat thionyl chloride (500 mL) was stirred for 2 h at 25°C. The excess thionyl chloride was distilled off in vacuo. The residuewas partitioned between saturated NaHCO₃ (400 mL) and dichloromethane(200 mL). The separated aqueous layer was extracted with dichloromethane(300 mL×3). The combined organic layers were dried, filtered andconcentrated in vacuo to give5-(chloromethyl)-2,2-difluorobenzo[d][1,3]dioxol (117.6 g, 89%), whichwas directly used in the next step.

2-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)acetonitrile

A mixture of 5-(chloromethyl)-2,2-difluorobenzo[d][1,3]dioxole (117.6 g,crude from last step) and NaCN (84 g, 1.7 mmol) in DMSO (800 mL) wasstirred for 2 h at 25° C. The reaction, mixture was poured into ice andextracted with EtOAc (500 mL×3). The combined organic layers were driedwith anhydrous Na₂SO₄ and concentrated in vacuo to give crude productwhich was purified by column chromatography (P.E./EtOAc 10:1) to give(2,2-difluorobenzo[d][1,3]dioxol-5-yl)methanol (77.8 g, 66%), ¹H NMR(300 MHz , CDCl₃) δ 7.06-7.07 (m, 3H), 3.75 (s, 2H).

1-(2,2-Difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid

1-(2,2-Difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid wasmade by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid, starting from2-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)acetonitrile. Yield (86%) of awhite solid. ¹H NMR (400 MHz, CDCl₃) δ 7.14-7.04 (m, 2H), 6.98-6.96 (m,1 H), 1.74-1.64 (m, 2H), 1.26-1.08 (m, 2H); ESI-MS m/z calc. 242.04,found 241.58 (M−1).

tert-Butyl thiazol-2-ylcarbamate

To a solution of aminothiazole (20.0 g, 199.7 mmol) and (Boc)₂O (48.0 g,219.7 mmol) in anhydrous THF (100 mL) were added DMAP (20 mg) and Et₃N(36.0 mL, 260.0 mmol). The reaction mixture was stirred at roomtemperature for 18 h, diluted with DCM and washed with 0.1 N HCl, H₂Oand brine. The organic layer was separated from the aqueous layer, driedover Na₂SO₄ and concentrated. The crude product was purified by columnchromatography (0-40% EtOAc/Hexane) to provide tert-butylthiazol-2-ylcarbamate as a white solid (20.7 g, 72%). ¹H-NMR (400 MHz,CDCl₃) δ 11.44 (s, 1H), 7.38 (d, J=3.6 Hz, 1H), 6.89 (d, J=3.6 Hz, 1H),1.58 (s, 9H); HPLC ret. time 2.61 min, 10-99% Ch₃CN, 5 min run; ESI-MS145.1 m/z (MH⁺).

tert-Butyl 4-methylthiazol-2-ylcarbamate

To a solution of 4-methylthiazol-2-amine (25 g, 219 mmol) and (Boc)₂O(53 g, 241 mmol) in anhydrous THF (110 mL) were added DMAP (250 mg) andEt₃N (39.6 mL, 285 mmol). The reaction mixture was stirred at roomtemperature for 18 h. Then the reaction was heated to reflux for 5 h,until no more starting material was detected by LC-MS. The reaction wascooled to room temperature and filtered to remove the precipitate. Thefiltrate was concentrated, then dissolved in CH₂Cl₂ and washed with 0.1N aqueous HCl, H₂O and brine. The organic layer was dried over MgSO₄ andconcentrated. The residue was suspended in hexane and then filtered toobtain tert-butyl 4-methylthiazol-2-ylcarbamate as a cream colored solid(30.9 g, 66%). ¹H-NMR (400 MHz, DMSO-d₆) δ 11.30 (s, 1H), 6.68 (d, J=1.0Hz, 1H), 2.20 (d, J=0.9 Hz, 3H), 1.47 (s, 9H); HPLC ret. time 2.68 min,10-99% CH₃CN, 5 min run; ESI-MS 215.3 m/z (MH⁺).

(R)-Dimethyl 2-hydroxysuccinate

To a solution of (R)-2-hydroxysuccinic acid (134 g, 1 mol) in CH₃OH (500mL) was added toluene-4-sulfonic acid (9.5 g, 0.05 mol). The mixture washeated to reflux overnight. Methanol was evaporated, and then water (250mL) was added to the residue. The mixture was basified with sat. NaHCO₃solution to pH 7-8 and extracted with ethyl acetate. The combinedorganic layers were washed with brine, dried over anhydrous Na₂SO₄, andconcentrated under reduced pressure to give (R)-dimethyl2-hydroxysuccinate (140 g, 86%). ¹H NMR (300 MHz, CDCl₃) δ 4.46 (dd, J=6Hz, 4.8 Hz, 1H), 3.74 (s, 3H), 3.64 (s, 3H), 3.42 (s, 1H), 2.69-2.85 (m,2H).

(R)-Methyl 3,4-dihydroxybutanoate

To a solution of (R)-dimethyl 2-hydroxysuccinate (140 g, 0.86 mol) inTHF (1400 mL) was added dropwise Me₂S.BH₃ (86 mL, 10 M) at 20° C. over30 min. The mixture was stilted at 20° C. for 30 min. NaBH₄ (1.63 g,42.9 mmol) was added at 10° C. and stirred at 10° C. for 30 min. Themixture was warmed to room temperature and stirred for 1 h. CH₃OH (200mL) was slowly added to the mixture while cooling in an ice-water bath.The resulting mixture was evaporated to give (R)-methyl3,4-dihydroxybutanoate (130 g, crude).

(R)-Methyl 4-chloro-3-hydroxybutanoate

To a solution of (R)-methyl 3,4-dihydroxybutanoate (125.1 g, 0.93 mol)in CH₂Cl₂ (1.8 L) was added PPh₃ (244.5 g, 0.93 mol) and slowly addedNCS (124.2 g, 0.93 mol) under ice water cooling. The mixture was stirredat 5° C. for 20 min and then stirred for 18 h at room temperature. Afterevaporating the solvent, the residue was purified by columnchromatography (P.E\E.A 20:1-5:1, gradient) to give (R)methyl4-chloro-3-hydroxybutanoate (33 g, 26% over 3 steps). ¹H NMR (300 MHz,CDCl₃)δ 4.18-4.25 (m, 1H), 3.68 (s, 3H), 3.55-3.60 (m, 2H), 3.32-3.33(d, J=42 Hz, 1H), 2.52-2.68 (m, 2H).

(R)-Methyl 3-(tert-butyldimethylsilyloxy)-4-chlorobutanoate

A solution of (R)-methyl 4-chloro-3-hydroxybutanoate (15 g, 98.7 mmol)in CH₂Cl₂ (240 mL) was stirred overnight withtert-butyl-chloro-dimethyl-silane (17.82 g, 118.2 mmol), imidazole (33.6g, 493.5 mmol) and a catalytic amount of DMAP (0.6 g, 4.92 mmol.) atroom temperature under N₂. The reaction mixture was poured into water(150 mL) and acidified to pH 6-7 by dropwise addition of cold aqueousHCl (0.5 M). The aqueous phase was extracted with CH₂Cl₂ (3×60 mL). Thecombined organic phases were washed with sat. Na₂CO₃ solution, brine,dried over anhydrous Na₂SO₄ and concentrated to give (R)-methyl3-(tert-butyldimethylsilyloxy)-4-chlorobutanoate (30 g, crude).

(R)-3-(tert-Butyldimethylsilyloxy)-4-chlorobutanal

To a solution of (R)-methyl3-(tert-butyldimethylsilyloxy)-4-chlorobutanoate (2.13 g, 104.7 mmol) inCH₃Cl₂ was added dropwise DIBAL-H (120 mL, 1 M in toluene, 120 mmol) at−78° C. under N₂ atmosphere. The mixture was stirred at −78° C. for 4 h.CH₃OH (80 mL) was added slowly to the reaction, mixture at −78° C. Thenthe temperature was warmed to room temperature gradually. The mixturewas filtered and the cake was washed with CH₂Cl₂. The combined filtrateswere concentrated under reduced pressure and purified by columnchromatography (PE) to give(R)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanal (13 g, 60% over 2steps). ¹H NMR (300 MHz, CDCl₃) δ 9.80 (t, J=1.6 Hz, 1H), 4.35-4.42 (m,1H), 3.53 (dd, J=11.1 Hz, 4.8 Hz, 1H), 3.46 (dd, J=12 Hz, 6.3 Hz, 1H),2.63-2.82 (m, 2H), 0.87 (s, 9H), 0.12 (s, 3H), 0.08 (s, 3H).

(S)-3-(tert-Butyldimethylsilyloxy)-4-chlorobutanal

(S)-3-(tert-Butyldimethylsilyloxy)-4-chlorobutanal was prepared by thesame route as (R)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanalstarting from (S)-2-hydroxysuccinic acid. ¹H NMR (400 MHz, CDCl₃) δ 9.72(dd, J=1.6, 2.2 Hz, 1H), 4.33-4.28 (m, 1H), 3.46 (dd, J=4.7, 11.1 Hz,1H), 3.39 (dd, J=6.4, 11.1 Hz, 1H), 2.68 (dd, J=1.5, 4.7 Hz, 1H), 2.62(dd, J=2.3, 6.8 Hz, 1H), 0.79 (s, 9H), 0.04 (s, 3H), 0.01 (s, 3H).

(R)-N-(2-Chlorobenzylidene)-2-methylpropane-2-sulfinamide

To a stirred anhydrous dioxane (500 mL) was added 2-chlorobenzaldehyde(34.8 g, 247.5 mmol) and the solution was cooled down to 0° C. in an icebath. A solution of (R)-2-methylpropanesulfinamide (30.0 g, 247.5 mmol)in anhydrous dioxane (100 mL) was added to the aldehyde solution.Ti(OPr)₄ (105.5 g, 371.3 mmol) was then slowly added to the solutionwhile stirring at 0° C. The reaction mixture was allowed to warm up toroom temperature and stirred for 18 h, then quenched with saturatedaqueous NaHCO₃ solution and filtered through a short plug of Celiteusing EtOAc. The organic layer was separated from the aqueous layer anddried over NaSO₄ and concentrated. The crude product was purified bycolumn chromatography (0-25% EtOAc/Hexane) to provide(R)-N-(2-chlorobenzylidene)-2-methylpropane-2-sulfinamide as a yellowliquid (45.1 g, 75%). ¹H-NMR (400 MHz, CDCl₃) δ 9.05 (s, 1H), 8.06 (dd,J=7.9, 1.1 Hz, 1H), 7.47-7.41 (m, 2H), 7.37-7.33 (m, 1H), 1.28 (s, 9H);HPLC ret. time 3.45 min, 10-99% CH₃CN, 5 min run; ESI-MS 244.3 m/z(MH⁺).

(R)-N-(3,4-Dichlorobenzylidene)-2-methylpropane-2-sulfinamide

(R)-N-(3,4-Dichlorobenzylidene)-2-methylpropane-2-sulfinamide was madeby the procedure used for(R)-N-(2-chlorobenzylidene)-2-methylpropane-2-sulfinamide, starting from3,4-dichlorobenzaldehyde and (R)-2-methylpropanesulfinamide. Yield (93%)of a yellow oil that crystallizes upon standing. ¹H-NMR (400 MHz, CDCl₃)□ 8.51 (s, 1H), 7.96 (d, J=1.9 Hz, 1H), 7.66 (dd, J=1.9, 8.3 Hz, 1H),7.57 (d, J=8.3 Hz, 1H), 1.27 (s, 9H); HPLC ret. time 3.72 min, 10-99%CH₃CN, 5 min run; ESI-MS 278.1 m/z (MH⁺).

(R)-N-(2-Chloro-4-fluorobenzylidene)-2-methylpropane-2-sulfinamide

(R)-N-(2-Chloro-4-fluorobenzylidene)-2-methylpropane-2-sulfinamide wasmade by the procedure used for(R)-N-(2-chlorobenzylidene)-2-methylpropane-2-sulfinamide, starting from2-chloro-4-fluorobenzaldehyde and (R)-2-methylpropanesulfinamide. Yield(74%) of a colorless solid. ¹H-NMR (400 MHz, CDCl₃) δ 9.00 (s, 1H), 8.11(dd, J=8.8, 6.2 Hz, 1H), 7.23 (dd, J=8.4, 2.5 Hz, 1H), 7.13-7.08 (m,1H), 1.29 (s, 9H); HPLC ret. time 3.46 min, 10-99% CH₃CN, 5 min run;ESI-MS 262.1 m/z (MH⁺).

(S)-N-(2-Chloro-4-fluorobenzylidene)-2-methylpropane-2-sulfinamide

(S)-N-(2-Chloro-4-fluorobenzylidene)-2-methylpropane-2-sulfinamide wasmade by the procedure used for(R)-N-(2-chlorobenzylidene)-2-methylpropane-2-sulfinamide, starting from2-chloro-4-fluorobenzaldehyde and (S)-2-methylpropanesulfinamide. Yield(78%) of a colorless solid. ¹H-NMR (400 MHz, CDCl₃) δ 9.00 (s, 1H), 8.11(dd, J=6.2, 8.8 Hz, 1H), 7.23 (dd, J=2.5, 8.4 Hz, 1H), 7.13-7.08 (m,1H), 1.29 (s, 9H); HPLC ret. time 3.49 min, 10-99% CH₃CN, 5 min run;ESI-MS 262.1 m/z MH⁺).

(R)-N-(4-Chloro-2-fluorobenzylidene)-2-methylpropane-2-sulfinamide

(R)-N-(4-Chloro-2-fluorobenzylidene)-2-methylpropane-2-sulfinamide wasmade by the procedure used for(R)-N-(2-chlorobenzylidene)-2-methylpropane-2-sulfinamide, starting from4-chloro-2-fluorobenzaldehyde and (R)-2-methylpropanesulfmamide. Yield(65%) of a colorless solid. ¹H-MMR (400 MHz, CDCl₃) δ 8.84 (s, 1H),7.97-7.93 (m, 1H), 7.28-7.19 (m, 2H), 1.27 (s, 9H); HPLC ret. time 3.53min, 10-99% CH₃CN, 5 min run; ESI-MS 262.0 m/z (MH⁺).

(R)-tert-Butyl5-((2-chloro-4-fluorophenyl)-(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamate

A solution of tert-butyl thiazol-2-ylcarbamate (2.0 g, 10.0 mmol) inanhydrous THF (25 mL) was stirred and cooled down to −78° C. To thissolution was slowly added n-BuLi (2.5 M in hexane: 8.0 mL, 20.0 mmol).Upon completion of n-BuLi addition, the mixture was maintained at −78°C. for 1 h. A solution of(R,E)-N-(2-chloro-4-fluorobenzylidiene)-2-methylpropane-2-sulfinamide(1.4 g, 5.4 mmol) in anhydrous THF (25 mL) previously cooled to −78° C.was slowly added to the above solution via canulation. The reaction waskept at −78° C. for 0.5 h, allowed to warm op to room temperature,quenched with aqueous NH₄Cl solution, and extracted with EtOAC (×3).The-combined organic layers were dried over MgSO₄ and concentrated. Thecrude product was purified by column chromatography (0-80% EtOAc/Hexane)to provide a diasteriomeric mixture of(R)-tert-butyl5-((2-chloro-4-fluorophenyl)(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamateas a yellow solid (2.1 g, 84%) that was used without furtherpurification.

(S)-tert-Butyl5-((2-chloro-4-fluorophenyl)-(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamate

(S)-tert-Butyl5-((2-chloro-4-fluorophenyl)-(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamatewas made by the procedure used, for (R)-tert-butyl5-((2-chloro-4-fluorophenyl)-(1,1dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamate,starting from tert-butyl thiazol-2-ylcarbamate and(S)-N-(2-chloro-4-fluorobenzylidene)-2-methylpropane-2-sulfinamide. Thecrude product was purified by column chromatography to provide adiasteriomeric mixture of (S)-tert-butyl5-((2-chloro-4-fluorophenyl)-(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamateas an orange-yellow solid (95%) that was used without furtherpurification.

(R)-tert-Butyl5-((4-chloro-2-fluorophenyl)-(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamate

(R)-tert-Butyl5-((4-chloro-2-fluorophenyl)-(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamatewas made by the procedure used for (R)-tert-butyl5-((2-chloro-4-fluorophenyl)-(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamate,starting from tert-butyl thiazol-2-ylcarbamate and(R)-N-(4-chloro-2-fluorobenzylidene)-2-methylpropane-2-sulfinamide. Thecrude product was purified by column chromatography to provide adiasteriomeric mixture of (R)-tert-butyl5-((4-chloro-2-fluorophenyl)-(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamate(78%) that was used without further purification,

(R)-tert-Butyl5-((3,4-dichlorophenyl)-(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamate

(R)-tert-Butyl5-((3,4-dichlorophenyl)-(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamatewas made by the procedure used for (R)-tert-butyl5-((2-chloro-4-fluorophenyl)-(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamate,starting from tert-butyl thiazol-2-ylcarbamate and(R)-N-(3,4-dichlorobenzylidene)-2-methylpropane-2-sulfinamide. The crudeproduct was purified by column chromatography (excess tert-butylthiazol-2-ylcarbamate is eluted at 70% EtOAc/hexane, the desired productfollows at 0-15% MeOH/EtOAc) to provide a diasteriomeric mixture of(R)-tert-Butyl5-((3,4-dichlorophenyl)-(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamateas a yellow solid (92%) that was used without further purification.

(R)-tert-Butyl5-((2-chlorophenyl)-(1,1-dimethylethylsulfinamido)methyl)-4-thiazol-2-ylcarbamate

(R)-tert-Butyl5-((2-chlorophenyl)-(1,1-dimethylethylsulfinamido)methyl)-4-thiazol-2-ylcarbamatewas made by the procedure used for (R)-tert-butyl5-((2-chloro-4-fluorophenyl)(1,1-dimethylethylsulfinamido)methyl)-4-thiazol-2-ylcarbamate,starting from tert-butyl 4-methylthiazol-2-ylcarbamate and(R)-N-(2-chlorobenzylidene)-2-methylpropane-2-sulfinamide. The crudeproduct was adsorbed onto silica gel and purified by columnchromatography (20-80% EtOAc/hexanes) to provide a diastereomericmixture of (R)-tert-butyl5-((2-chlorophenyl)-(1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-ylcarbamateas a cream colored solid (90%) that was used without furtherpurification.

(R)-N-((S)-(2-Aminothiazol-5-yl)(2-chloro-4-fluorophenyl)methyl)-2-methylpropane-2-sulfinamide

To a solution of of (R)-tert-Butyl5-((2-chloro-4-fluorophenyl)(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamate(1.6 g, 3.5 mmol) in CH₂Cl₂ (6 mL) was added TFA (6 mL). The reactionwas stirred at room temperature for 2 h. The TFA and CH₂Cl₂ were removedunder vacuum. The crude product was re-dissolved in CH₂Cl₂, washed withaqueous NaHCO₃ solution, dried over MgSO₄ and concentrated. The crudeproduct was recrystallized from EtOAc to provide(R)-N-((S)-(2-aminothiazol-5-yl))(2-chloro-4-fluorophenyl)methyl)2-methylpropane-2-sulfinamideas colorless solid (672 mg, 53%), ¹H NMR (400 MHz, CDCl₃) δ 7.56 (dd,J=8.7, 5.9 Hz, 1H), 7.16 (dd, J=8.4, 2.6 Hz, 1H), 7.06 (td, J=8.2, 2.6Hz, 1H), 6.89 (d, J=0.8 Hz, 1H), 6.07 (d, J=4.6 Hz, 1H), 5.22 (d, J=2.9Hz, 2H), 4.06 (d, J=4.6 Hz, 1H), 1.28 (s, 9H); HPLC ret. time 2.28 min,10-99% CH₃CN, 5 min run; ESI-MS 362.3 m/z (MH⁺).

(S)-N-((R)-(2-Aminothiazol-5-yl)(2-chloro-4-fluorophenyl)methyl)-2-methylpropane-2-sulfinamide

(S)-N-((R)-(2-Aminothiazol-5-yl)(2-chloro-4-fluorophenyl)methyl)-2-methylpropane-2-sulfinamidewas made by the procedure used for(R)-N-((R)-(2-aminothiazol-5-yl)(2-chloro-4-fluorophenyl)methyl)-2-methylpropane-2-sulfinamide,starting from (S)-tert-Butyl5-((4-chloro-2-fluorophenyl)(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamate.The crude product was purified by column chromatography (0-5%EtOH/EtOAc) to provide the desired product (38%). ¹H-NMR (400 MHz,CDCl₃) δ 9.14 (br s, 2H), 7.54 (dd, J=5.7, 8.7 Hz, 1H), 7.21 (dd, J=2.6,8.1 Hz, 1H), 7.13-7.08 (m, 1H), 6.73 (d, J=0.6 Hz, 1H), 6.00 (d, J=5.6Hz, 1H), 4.67 (d, J=5.7 Hz, 1H), 1.26 (s, 9H); HPLC ret. time 2.23 min,10-99% CH₃CN, 5 min run; ESI-MS 362.3 m/z (MH⁺).

(R)-N-((S)-(2-Aminothiazol-5-yl)(2-chloro-4-fluorophenyl)methyl)-2-methylpropane-2-sulfinamide

(R)-N-((S)-(2-Aminothiazol-5-yl)(2-chloro-4-fluorophenyl)methyl)-2-methylpropane-2-sulfinamidewas made by the procedure used for(R)-N-((S)-(2-aminothiazol-5-yl)(2-chloro-4-fluorophenyl)methyl)-2-methylpropane-2-sulfinamide,starting from (R)-tert-butyl5-((2-chloro-4-fluorophenyl)(1,1-dimethylethylsulfinamido)methyl)thiazol-2ylcarbamate.The crude product was purified by column chromatography (0-5%EtOH/EtOAc) to provide an orange solid (87%). ¹H-NMR (400 MHz, CDCl₃) δ8.62 (br s, 2H), 7.36 (t, J=8.1 Hz, 1H), 7.21 (dd, J=1.7, 8.3 Hz, 1H),7.16 (dd, J=1.9, 10.1 Hz, 1H), 6.74 (s, 1H), 5.81 (d, J=5.9 Hz, 1H),4.18 (d, J=5.9 Hz, 1H), 1.25 (s, 9H); HPLC ret. time 2.29 min, 10-99%Ch₃CN, 5 min run; ESI-MS 362.3 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide

To a solution of 1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid(618 mg, 3.0 mmol) in anhydrous CH₂C₂ (6 mL) was slowly added (COCl)₂(0.3 mL, 3.4 mmol) at −10° C. followed by DMF (3 drops). The reactionmixture was stirred at −10° C. for 0.5 h. The excess (COCl)₂ was removedunder vacuum. The acid chloride (10.5 mmol) was then dissolved inanhydrous CH₂Cl₂ (3 mL) and was slowly added to a solution of(R)-N-((S)-(2-aminothiazol-5-yl)(2-chloro-4-fluorophenyl)methyl)-2-methylpropane-2-sulfinamide(648 mg, 1.8 mmol) and Et₃N (1.8 mL, 6 mmol) in anhydrous CH₂Cl₂ (3 mL).The reaction mixture was stirred at room temperature for 1 h, dilutedwith CH₂Cl₂ and washed with 1N HCl, NaHCO₃ and brine. The organic layerwas separated from the aqueous layer and dried over MgSO₄ andconcentrated. The crude product was purified by column chromatography(40-60% EtOAc/Hexane) to provide1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamideas a colorless solid (680 mg, 69%). ¹H-NMR (400 MHz, CDCl₃) δ 8.60 (s,1H), 7.60 (dd, J=8.7, 5.9 Hz, 1H), 7.28 (d, J=2.0 Hz, 1H), 7.13 (dd,J=8.3, 2.6 Hz, 1H), 7.05 (td, J=8.2, 2.6 Hz, 1H), 6.90 (td, J=8.3, 1.7Hz, 2H), 6.83 (d, J=7.9 Hz, 1H), 6.18 (d, J=4.3 Hz, 1H), 6.04 (s, 2H),3.90 (d, J=4.3 Hz, 1H), 1.73 (td, J=5.4, 2.0 Hz, 2H), 1.28 (t, J=7.1 Hz,2H), 1.29 (s, 9H). HPLC ret. time 3.65 min, 10-99% CH₃CN, 5 min run;ESI-MS 550.5 m/z (MH⁺).

N-(5-((S)-(2-Chloro-4-fluorophyenyl)((R)-1,1-dimethyethylsulfinamido)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide

N-(5-((S)-(2-Chloro-4-fluorophyenyl)((R)-1,1-dimethyethylsulfinamido)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamidewas made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide,starting from1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid and(R)-N-((S)-(2-aminothiazol-5-yl)(2-chloro-4-fluorophenyl)methyl)-2-methylpropane-2-sulfinamide.Yield (77%). ¹H-NMR (400 MHz, CDCl₃) δ 8.63 (s, 1H), 7.59 (dd, J=8.7,5.9 Hz, 1H), 7.28 (s, 1H), 7.21-7.11 (m, 4H), 7.06 (td, J=8.2, 2.5 Hz,1H), 6.18 (d, J=4.2 Hz, 1H), 3.92 (d, J=4.3 Hz, 1H), 1.82-1.79 (m, 2H),1.32-1.25 (m, 11H); HPLC ret. time 3.90 min, 10-99% CH₃CN, 5 min run;ESI-MS 586.3 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(2-chloro-4-fluorophenyl)((S)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(2-chloro-4-fluorophenyl)((S)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamidewas made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamidefrom 1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid and(S)-N-((R)-(2-aminothiazol-5-yl)(2-chloro-4-fluorophenyl)methyl)-2-methylpropane-2-sulfinamide.Yield (50%). ¹H-NMR (400 MHz, DMSO-d₆) δ 11.10 (s, 1H), 7.78 (dd, J=6.3,8.8 Hz, 1H), 7.45 (dd, J=2.6, 8.8 Hz, 1H), 7.35 (td, J=8.6, 2.7 Hz, 1H),7.03 (s, 1H), 6.95 (s, 1H), 6.86 (s, 2H), 6.45 (d, J=6.7 Hz, 1H), 6.00(s, 2H), 5.95 (d, J=6.5 Hz, 1H), 1.46-1.43 (m, 2H), 1.25-1.24 (m, 2H),1.14 (s, 9H); HPLC ret. time 3.65 min, 10-99% CH₃CN, 5 min run; ESI-MS550.5 m/z (MH⁺).

N-(5-((R)-(2-Chloro-4-fluorophenyl)((S)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide

N-(5-((R)-(2-Chloro-4-fluorophenyl)((S)-1,1-dimethyethylsulfinamido)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamidewas made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide,starting from1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid and(S)-N-((R)-(2-aminothiazol-5-yl)(2-chloro-4-fluorophenyl)methyl)-2-methylpropane-2-sulfinamide.Yield (50%). ¹H-NMR (400 MHz, DMSO-d₆) δ 11.38 (s, 1H), 7.78 (dd, J=0.2,8.8 Hz, 1H), 7.47-7.44 (m, 2H), 7.37-7.32 (m, 2H), 7.21 (dd, J=1.6, 8.3Hz, 1H), 7.04 (s, 1H), 6.45 (d, J=6.6 Hz, 1H), 5.95 (d, J=6.5 Hz, 1H),1.54-1.50 (m, 2H), 1.22-1.21 (m, 2H), 1.14 (s, 9H); HPLC ret. time 3.90min, 10-99% Ch₃CH, 5 min run; ESI-MS 586.3 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(4-chloro-2-fluorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(4-chloro-2-fluorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamidewas made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamidestarting from 1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acidand(R)-N-((S)-(2-aminothiazol-5-yl)(4-chloro-2-fluorophenyl)methyl)-2-methylpropane-2-sulfinamide.Yield (64%). ¹H-NMR (400 MHz, CDCl₃) δ 8.56 (s, 1H), 7.43 (t, J=8.1 Hz,1H), 7.18 (s, 1H), 7.16 (dd, J=1.5, 8.3 Hz, 1H), 7.08 (dd, J=2.0, 10.0Hz, 1H), 6.89 (td, J=8.0, 1.7 Hz, 2H), 6.81 (d, J=7.9 Hz, 1H), 6.01-5.99(m, 3H), 3.90 (d, J=5.0 Hz, 1H), 1.73-1.70 (m, 2H), 1.28-1.23 (m, 11B);. HPLC ret. time 3.69 min, 10-99% CH₃CN, 5 min run; ESI-MS 550.5 m/z(MH⁺).

N-(5-((S)-(4-Chloro-2-fluorophyenyl)((R)-1,1-dimethyethylsulfinamido)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide

N-(5-((S)-(4-Chloro-2-fluorophyenyl)((R)-1,1-dimethyethylsulfinamido)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamidewas made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide,starting from1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid and(R)-N-((S)-(2-aminothiazol-5-yl)(4-chloro-2-fluorophenyl)methyl)-2-methylpropane-2-sulfinamide.Yield (53%), ¹H-NMR (400 MHz, DMSO-d₆) δ 11.37 (s, 1H), 7.66 (t, J=8.3Hz, 1H), 7.45-7.42 (m, 2H), 7.38 (dd, J=2.0, 8.4 Hz, 1H), 7.34 (d, J=8.3Hz, 1H), 7.21 (dd, J=1.5, 8.3 Hz, 1H), 7.11 (s, 1H), 6.33 (d, J=6.7 Hz,1H), 5.89 (d, J=6.5 Hz, 1H), 1.55-1.52 (m, 2H), 1.24-1.22 (m, 2H), 1.13(d, J=9.5 Hz, 9H); HPLC ret. time 3.93 min, 10-99% CH₃CN, 5 min run;ESI-MS 586,5 m/z (MH⁺).

(S)-N-(5-(Amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

To a solution, of1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide(659 mg, 1.2 mmol) in MeOH (5 mL) was added 4M HCl in dioxane (1.8 mL,7.2 mmol). The reaction mixture was stirred at room temperature for 1.5h and evaporated to dryness. The crude product was dissolved in CH₂Cl₂.The organic layer was washed with aqueous NaHCO₃ solution (50 mL×2),brine (50 mL×1), dried over MgSO₄ and concentrated to provide(S)-N-(5-(Amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideas a colorless solid that was used without further purification. ¹H-NMR(400 MHz, CDCl₃) δ 8.48 (br s, 1H), 7.63 (dd, J=6.1, 8.7 Hz, 1H), 7.18(s, 1H), 7.08 (dd, J=2.6, 8.4 Hz, 1H), 6.99 (td, J=8.3, 2.6 Hz, 1H),6.91-6.86 (m, 2H), 6.81 (d, J=7.9 Hz, 1H), 6.01 (s, 2H), 5.78 (s, 1H),3.80 (s, 2H), 1.74-1.67 (m, 2H), 1.30-1.22 (m, 2H); HPLC ret. time 2.67min, 10-99% CH₃CN, 5 min run: ESI-MS 446.3 m/z (MH⁺).

(S)-N-(5-(Amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

(S)-N-(5-(Amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamidewas made by the procedure used for(S)-N-(5-(amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide, starting fromN-(5-((S)-(2-chloro-4-fluorophyenyl)((R)-1,1-dimethyethylsulfinamido)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide.The crude product was used without further purification. ¹H-NMR (400MHz, CDCl₃) δ 8.35 (s, 1H), 7.65 (dd, J=6.1, 8.7 Hz, 1H), 7.22-7.17 (m,3H), 7.12-7.10 (m, 2H), 7.02 (td, J=8.3, 2.6 Hz, 1H), 5.80 (s, 1H), 3.83(s, 2H), 1.83-1.77 (m, 2H), 1.28-1.25 (m, 2H); HPLC ret. time 2.87 min,10-99% CH₃CN, 5 min run; ESI-MS 482.3 m/z (MH⁺).

(R)-N-(5-(Amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

(R)-N-(5-(Amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamidewas made by the procedure used for(S)-N-(5-(amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamidestarting from1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(2-chloro-4-fluorophenyl)((S)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide.The crude product was purified by column chromatography (0-2.5%Et₃N/EtOAc) to provide a pale yellow solid (77%). ¹H-NMR (400 MHz,DMSO-d₆) δ 10.80 (br s, 1H), 7.80 (dd, J=6.4, 8.8 Hz, 1H), 7.38 (dd,J=2.6, 8.8 Hz, 1H), 7.27 (td, J=8.5, 2.7 Hz, 1H), 7.07 (d, J=0.9 Hz,1H), 6.95 (s, 1H), 6.86 (d, J=0.9 Hz, 2H), 6.00 (s, 2H), 5.53 (s, 1H),1.44-1.41 (m, 2H), 1.13-1.10 (m, 2H); HPLC ret. time 2.67 min, 10-99%CH₃CN, 5 min run; ESI-MS 446.3 m/z (MH⁺).

(R)-N-(5-(Amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

(R)-N-(5-(Amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamidewas made by the procedure used for(R)-N-(5-(amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide, starting fromN-(5-((R)-(2-chloro-4-fluorophenyl)((S)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(79%). ¹H-NMR (400 MHz, DMSO-d₆) δ 7.80 (dd, J=6.4, 8.8 Hz, 1H), 7.43(d, J=1.7 Hz, 1H), 7.38 (dd, J=2.6, 8.8 Hz, 1H), 7.34 (d, J=8.3 Hz, 1H),7.27 (td, J=8.5, 2.6 Hz, 1H), 7.20 (dd, J=1.8, 8.3 Hz, 1H), 7.07 (d,J=0.9 Hz, 1H), 5.53 (s, 1H), 1.52-1.49 (m, 2H), 1.21-1.19 (m, 2H); HPLCret. time 2.91 min, 10-99% CH₃CN, 5 min run; ESI-MS 482 m/z (MH⁺).

(S)-N-(5-(Amino(4-chloro-2-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

(S)-N-(5-(Amino(4-chloro-2-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamidewas made by the procedure used for(R)-N-(5-(amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide,starting from1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(4-chloro-2-fluorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide(79%). ¹H NMR (400 MHz, CDCl₃) δ 8.48 (s, 1H), 7.47 (t, J=8.2 Hz, 1H),7.14-7.12 (m, 2H), 7.05 (dd, J=2.0, 10.1 Hz, 1H), 6.91-6.87 (m, 2H),6.81 (d, J=7.9 Hz, 1H), 6.01 (s, 2H), 5.61 (s, 1H), 1.86 (s, 2H), 1.71(dd, J=3.6, 6.7 Hz, 2H), 1.22-1.19 (m, 2H); HPLC ret. time 2.71 min,10-99% CH₃CN, 5 min run; ESI-MS 446.3 m/z (MH⁺).

(S)-N-(5-(Amino(4-chloro-2-fluorophenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

(S)-N-(5-(Amino(4-chloro-2-fluorophenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamidewas made by the procedure used for(R)-N-(5-(amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide,starting fromN-(5-((S)-(4-chloro-2-fluorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(88%). ¹H NMR (400 MHz, DMSO-d₆) δ 11.20 (br s, 1H), 7.66 (t, J=8.3 Hz,1H), 7.43 (d, J=1.7 Hz, 1H), 7.39-7.30 (m, 3H), 7.20 (dd, J=1.7, 8.3 Hz,1H), 7.09 (s, 1H), 5.45 (s, 1H), 1.52-1.49 (m, 2H), 1.22-1.20 (m, 2H);HPLC ret. time 2.94 min, 10-99% CH₃CN, 5 min run; ESI-MS 482.3 m/z(MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide

To a solution of(S)-N-(5-(amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(1.2 mmol) in MeOH (10 mL) was added(R)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanal (341 mg, 1.4 mmol).The reaction mixture was stirred at room temperature for 5 min. ThenNaBH₄ (68 mg, 1.8 mmol) was added and stirring was continued at roomtemperature for 1 h. The reaction was diluted with H₂O and extractedwith EtOAc. The combined organic layers were washed with brine and driedover MgSO₄. After the removal of solvent, the residue was purified bycolumn chromatography (10-20% EtOAc/Hexane) to afford1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide(446 mg, 59% over two steps). ¹H NMR (400 MHz, CDCl₃) δ 8.45 (s, 1H),7.79 (dd, J=8.6, 6.3 Hz, 1H), 7.25 (d, J=6.4 Hz, 1H), 7.01-6.94 (m, 2H),6.85 (td, J=8.3, 1.7 Hz, 2H), 6.78 (d, J=7.9 Hz, 1H), 5.99 (s, 2H), 4.99(s, 1H), 4.35-4.30 (m, 1H), 2.77 (dd, J=9.9, 6.2 Hz, 1H), 2.51-2.48 (m,2H), 2.36 (dd, J=9.9, 4.3 Hz, 1H), 2.06-2.01 (m, 1H), 1.71-1.65 (m, 3H),1.18 (t, J=3.0 Hz, 2H), 0.85 (s, 9H), −0.01 (d, J=7.3 Hz, 6H); HPLC ret.time 3.59 min, 10-99% CH₃CN, 5 min run; ESI-MS 630.6 m/z (MH⁺).

N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1yl)(2-chloro-4-fluorophyenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide

N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1yl)(2-chloro-4-fluorophyenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamidewas made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide,starting from(S)-N-(5-(amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(2,2-fluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideand (R)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanal (56% over twosteps). ¹H NMR (400 MHz, CDCl₃) δ 8.31 (s, 1H), 7.79 (dd, J=8.6, 6.3 Hz,1H), 7.25 (d, J=7.9 Hz, 1H), 7.13 (td, J=8.4, 1.6 Hz, 2H), 7.06 (d,J=8.1 Hz, 1H), 7.01-6.95 (m, 2H), 4.99 (s, 1H), 4.35-4.30 (m, 1H), 2.76(dd, J=9.8, 6.2 Hz, 1H), 2.50 (t, J=6.9 Hz, 2H), 2.37 (dd, J=10.0, 4.2Hz, 1H), 2.08-1.98 (m, 1H), 1.79-1.64 (m, 3H), 1.24-1.19 (m, 2H), 0.85(s, 9H), −0.01 (d, J=7.4 Hz, 6H); HPLC ret. time 3.80 min, 10-99% CH₃CN,5 min run; ESI-MS 666.4 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamidewas made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide,starting from of(R)-N-(5-(amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideand (R)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanal (99%). ¹H NMR(400 MHz, DMSO-d₆) δ 11.01 (s, 1H), 7.77 (dd, J=6.4, 8.8 Hz, 1H),7.38-7.36 (m, 2H), 7.29 (td, J=8.5, 2.7 Hz, 1H), 6.94 (s, 1H), 6.85 (m,2H), 5.99 (s, 2H), 4.99 (s, 1H), 4.38-4.34 (m, 1H), 2.83 (dd, J=6.2, 9.9Hz, 1H), 2.58-2.54 (m, 1H), 2.46-2.42 (m, 1H), 2.14 (dd, J=3.7, 9.9 Hz,1H), 2.05-1.99 (m, 1H), 1.68-1.57 (m, 1H), 1.43-1.40 (m, 2H), 1.12-1.10(m, 2H), 0.82 (s, 9H), −0.00 (s, 3H), −0.02 (s, 3H); HPLC ret. time 3.60min, 10-99% CH₃CN, 5 min run; ESI-MS 630.5 m/z (MH⁺).

N-(5-((R)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1yl)(2-chloro-4-fluorophyenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide

N-(5-((R)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1yl)(2-chloro-4-fluorophyenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamidewas made by the procedure-used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide,starting from(R)-N-(5-(amino(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideand (R)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanal (92%). ¹H NMR(400 MHz, DMSO-d₆) δ 11.32 (s, 1H), 7.77 (dd, J=6.4, 8.8 Hz, 1H), 7.43(d, J=1.4 Hz, 1H), 7.39-7.30 (m, 4H), 7.20 (dd, J=1.5, 8.3 Hz, 1H), 4.99(s, 1H), 4.40-4.35 (m, 1H), 2.83 (dd, J=6.1, 9.9 Hz, 1H), 2.60-2.52 (m,1H), 2.46-2.41 (m, 1H), 2.15 (dd, J=3.7, 9.9 Hz, 1H), 2.07-1.99 (m, 1H),1.63-1.57 (m, 1H), 1.51-1.48 (m, 2H), 1.25-1.24 (m, 2H), 0.83 (s, 9H),0.01 (s, 3H), −0.02 (s, 3H); HPLC ret. time 3.79 min, 10-99% CH₃CN, 5min run; ESI-MS 666.3 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(4-chloro-2-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(4-chloro-2-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamidewas made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamidestarting from of(S)-N-(5-(amino(4-chloro-2-fluorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideand (R)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanal (89%). ¹H NMR(400 MHz, CDCl₃) δ 8.48 (s, 1H), 7.64 (t, J=8.1 Hz, 1H), 7.22 (s, 1H),7.11 (dd, J=1.9, 8.4 Hz, 1H), 6.99 (dd, J=2.0, 9.9 Hz, 1H), 6.90-6.85(m, 2H), 6.80 (d, J=7.9 Hz, 1H), 6.00 (s, 2H), 4.91 (s, 1H), 4.35-4.33(m, 1H), 2.83 (dd, J=6.2, 9.8 Hz, 1H), 2.55-2.50 (m, 2H), 2.35 (dd,J=4.4, 9.8 Hz, 1H), 2.07-2.02 (m, 1H), 1.93-1.91 (m, 1H), 1.73-1.70 (m,2H), 1.22-1.20 (m, 2H), 0.86 (s, 9H), 0.01 (s, 3H), 0.00 (s, 3H); HPLCret. time 3.59 min, 10-99% CH₃CN, 5 min run; ESI-MS 630.5 m/z (MH⁺).

N-(5-((S)-((R)-3-(tert-Butyldimethylsilyloxy)pyrrolidin-1yl)(4-chloro-2-fluorophyenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide

N-(5-((S)-((R)-3-(tert-Butyldimethylsilyloxy)pyrrolidin-1yl)(4-chloro-2-fluorophyenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamidewas made by the procedure used, for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide,starting from(S)-N-(5-(amino(4-chloro-2-fluorophenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideand (R)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanal (90%). ¹H NMR(400 MHz, DMSO-d₆) δ 11.33 (s, 1H), 7.63 (t, J=8.2 Hz, 1H), 7.43-7.32(m, 5H), 7.20 (dd, J=1.5, 8.3 Hz, 1H), 4.93 (s, 1H), 4.39-4.34 (m, 1H),2.69 (dd, J=6.1, 9.8 Hz, 1H), 2.49-2.45 (m, 2H), 2.34 (dd, J=3.9, 9.9Hz, 1H), 2.10-1.99 (m, 1H), 1.62-1.55 (m, 1H), 1.52-1.49 (m, 2H),1.24-1.19 (m, 2H), 0.83 (s, 9H), 0.01 (s, 3H), −0.01 (s, 3H); HPLC ret.time 3.79 min, 10-99% CH₃CN, 5 min run; ESI-MS 666.3 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide

A mixture of1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide(365 mg, 0.58 mmol) and TBAF (1M in THF, 2.3 mL, 2.3 mmol) was stirredat room temperature overnight The reaction was diluted with H₂O andextracted with EtOAc. The combined organic layers were washed with brineand dried over MgSO₄. After the removal of solvent, the residue waspurified by column chromatography (20-50% EtOAc/Hexane) to afford1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide(183 mg, 61%, >99% de). ¹H-NMR (400 MHz, DMSO-d₆) δ 8.41 (s, 1H), 7.72(t, J−7.1 Hz, 1H), 7.20 (d, J=83 Hz, 2H), 6.95 (t, J=7.2 Hz, 2H), 6.80(td, J=8.5, 1.6 Hz, 2H), 6.73 (d, J=7.9 Hz, 1H), 5.94 (s, 2H), 4.95 (s,1H), 4.25 (s, 1H), 2.75 (s, 1H), 2.58-2.56 (m, 1H), 2.42 (s, 1H),2.28-2.27 (m, 1H), 2.12-2.11 (m, 1H), 1.71-1.61 (m, 3H), 1.19-1.10 (m,20); HPLC ret. time 2.68 min, 10-99% CH₃CN, 5 min run; ESI-MS 516.2 m/z(MH⁺).

N-(5-((S)-(2-Chloro-4-fluorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide

N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamidewas made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide,starting fromN-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(77%). ¹H NMR (400 MHz, DMSO-d₆) δ 8.36 (s, 1H), 7.80 (t, J=7.2 Hz, 1H),7.30 (d, J=10.6 Hz, 1H), 7.17 (td, J=8.5, 1.7 Hz, 2H), 7.11-7.01 (m,4H), 5.05 (s, 1H), 4.35-4.33 (m, 1H), 2.84-2.83 (m, 1H), 2.67-2.64 (m,1H), 2.50 (dd, J=10.0, 4.9 Hz, 1H), 2.38-2.34 (m, 1H), 2.20 (dd, J=12.9,5.8 Hz, 1H), 1.87-1.76 (m, 3H), 1.30-1.23 (m, 2H); HPLC ret. time 2.91min, 10-99% CH₃CN, 5 min run; ESI-MS 55.2.4 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(2-chloro-4-fluorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(2-chloro-4-fluorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamidewas made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide,starting from1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide(61%, >99% de). ¹H NMR (400 MHz, DMSO-d₆) δ 11.01 (s, 1H), 7.80 (dd,J=6.4, 8.8 Hz, 1H), 7.39-7.36 (m, 2H), 7.30 (td, J=8.5, 2.6 Hz, 1H),6.95 (s, 1H), 6.85 (s, 2H), 6.00 (s, 2H), 4.94 (s, 1H), 4.72 (d, J=4.3Hz, 1H), 4.19-4.18 (m, 1H), 2.70-2.66 (m, 1H), 2.60-2.56 (m, 1H),2.34-2.30 (m, 1H), 2.20 (dd, J=3.1, 9.9 Hz, 1H), 2.00-1.95 (m, 1H),1.60-1.57 (m, 1H), 1.44-1.41 (m, 2H), 1.14-1.08 (m, 2H); HPLC ret. time2.72 min, 10-99% CH₃CN, 5 min run; ESI-MS 516.3 m/z (MH⁺).

N-(5-((R)-(2-Chloro-4-fluorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

N-(5-((R)-(2-Chloro-4-fluorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamidewas made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide,starting fromN-(5-((R)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(2-chloro-4-fluorophenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(73%, >99% de). ¹H NMR (409 MHz, DMSO-d₆) δ 11.32 (s, 1H), 7.80 (dd,J=6.4, 8.8 Hz, 1H), 7.43 (d, J=1.4 Hz, 1H), 7.39-7.28 (m, 4H), 7.20 (dd,J=1.6, 8.3 Hz, 1H), 4.94 (s, 1H), 4.72 (d, J=4.3 Hz, 1H), 4.20-4.19 (m,1H), 2,71-2.67 (ms 1H, 2.63-2.57 (m, 1H), 2.34-2.29 (m, 1H), 2.20 (dd,J=3.2, 9.8 Hz, 1H), 2.02-1.93 (m, 1H), 1.62-1.54 (m, 1H), 1.54-1.48 (m,2H), 1.23-1.17 (m, 2H); HPLC ret. time 2.94 min, 10-99% CH₃CNS 5 minrun; ESI-MS 552.5 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(4-chloro-2-fluorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(4-chloro-2-fluorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamidewas made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide,starting from1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(4-chloro-2-fluorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide(37%, >99% de). ¹H NMR (400 MHz, CDCl₃) δ 8.48 (s, 1H), 7.62 (t, J=8.1Hz, 1H), 7.22 (s, 1H), 7.13 (dd, J=1.9, 8.4 Hz, 1H), 7.01 (dd, J=2.1,9.9 Hz, 1H), 6.90-6.85 (m, 2H), 6.81 (d, J=7.9 Hz, 1H), 6.01 (s, 2H),4.92 (s, 1H), 4.34-4.28 (m, 1H), 2.86-2.80 (m, 1H), 2.63 (d,J=10.1 Hz,1H), 2.50-2.46 (m, 1H), 2.34-2.23 (m, 1H), 2.21-2.12 (m, 1H), 1.80-1.70(m, 4H), 1.24-1.18 (m, 2H); HPLC ret. time 2.77 min, 10-99% CH₃CN, 5 minrun; ESI-MS 516.3 m/z (MH⁺).

N-(5-((S)-(4-chloro-2-fluorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

N-(5-((S)-(4-chloro-2-fluorophenyl)((R)-3-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamidewas made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chloro-4-fluorophenyl)((R)-hydroxypyrrolidin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide,starting fromN-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolidin-1-yl)(4-chloro-2-fluorophenyl)methyl)thiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(70%, >99% de). ¹H NMR (400 MHz, DMSO-d₆) δ 11.32 (s, 1H), 7.66 (t,J=8.1 Hz, 1H), 7.43 (d, J=1.5 Hz, 1H), 7.39-7.33 (m, 4H), 7.20 (dd,J=1.6, 8.3 Hz, 1H), 4.88 (s, 1H), 4.75 (d, J=4.4 Hz, 1H), 4.21-4.16 (m,1H), 2.57 (dd, J=6.1, 9.8 Hz, 1H), 2.51-2.44 (m, 1H), 2.41-2.33 (m, 2H),2.04-1.95 (m, 1H), 1.60-1.45 (m, 3H), 1.24-1.15 (m, 2H); HPLC ret. time2.97 min, 10-99% Ch₃CN, 5 min run; ESI-MS 552.5 m/z (MH⁺).

(R)-N-((R)-(2-Amino-4-methylthiazol-5-yl)(2-chlorophenyl)methyl)-2-methylpropane-2-sulfinamideand(R)-N-((S)-(2-Amino-4-methylthiazol-5-yl)(2-chlorophenyl)methyl)-2-methylpropane-2-sulfinamide

To a solution of of (R)-tert-Butyl5-((2-chlorophenyl)(1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-ylcarbamate,(19.0 g, 41.6 mmol) in CH₂Cl₃ (83 mL) was added TFA (83 mL). Thereaction was stirred at room temperature for 1 h. The reaction wasconcentrated and then partitioned between CH₂Cl₂ and saturated aqueousNaHCO₃ solution. The aqueous layer was basirled to pH>12 by adding 1NNaOH solution and extracted with Ch₂Cl₂. The combined organic layerswere dried over MgSO₄ and concentrated. The crude product was adsorbedonto silica gel and purified by column chromatography (70-100%EtOAc/hexanes. The EtOAc contained 2% NH₄OH and was stirred to maintainthe mixture) to provide(R)-N-((R)-(2-amino-4-methylthiazol-5-yl)(2-chlorophenyl)methyl)-2-methylpropane-2-sulfinamideand(R)-N-((S)-(2-amino-4-methylthiazol-5-yl)(2-chlorophenyl)methyl)-2-methylpropane-2-sulfinamideand a mixture of both diastereomers (7.1 g, 47%). First eluted product(Isomer A) (white solid, 3.0 g, 20%, >99% de); ¹H NMR (400 MHz, CDCl₃) δ7.60 (dd, J=1.6, 7.7 Hz, 1H), 7.38-7.22 (m, 3H), 6.15 (d, J=2.5 Hz, 1H),4.94 (s, 2H), 3.87 (d, J=2.3 Hz, 1H), 2.34 (s, 3H), 1.27 (s, 9H); HPLCret. time 3.08 min, 10-99% CH₃CN, 15 min run; ESI-MS 358.3 m/z (MH⁺).Second eluted product (Isomer B) (yellow waxy solid, 5.0 g, 33% 98% de);¹H NMR (400 MHz, CDCl₃) δ 7.64 (dd, J=7.7, 1.7 Hz, 1H), 7.36 (dd, J=7.8,1.4 Hz, 1H), 7.32-7.21 (m, 2H), 6.19 (d, J=1.8 Hz, 1H), 4.86 (s, 2H),3.65 (d, J=1.2 Hz, 1H), 2.33 (s, 3H), 1.26 (s, 9H); HPLC ret. time 3.77min, 10-99% CH₃CN, 15 min run; ESI-MS 358.3 m/z (MH⁺).

(R)-N-((R)-(2-aminothiazol-5-yl)(3,4-dichlorophenyl)methyl)-2-methylpropane-2-sulfinamideand(R)-N-((S)-(2-aminothiazol-5-yl)(3,4-dichlorophenyl)methyl)-2-methylpropane-2-sulfinamide

(R)-N-(S)-1-(2-Aminothiazol-5-yl)-1-(3,4-dichlorophenyl)methyl)-1,1-dimethylethylsulfinamideand(R)-N-(R)-1-(2-aminothiazol-5-yl)(3,4-dichlorophenyl)methyl)-1,1-dimethylethylsulfinamidewere made by the procedure for((R)-(2-amino-4-methylthiazol-5-yl)(2-chlorophenyl)methyl)-2-methylpropane-2-sulfinamideand(R)-N-((S)-(2-amino-4-methylthiazol-5-yl)(2-chlorophenyl)methyl)-2-methylpropane-2-sulfinamidestarting from (R)-tert-butyl5-((3,4-dichlorophenyl)(1,1-dimethylethylsulfinamido)methyl)thiazol-2-ylcarbamate.The crude product was purified by column chromatography (5% EtOH/EtOAcelutes Isomer A, 20% EtOH/EtOAc elutes Isomer B). Isomer A (yellowsolid, 5.67 g, 35%, >99% de); ¹H NMR (400 MHz, CDCl₃) δ 7.40 (d, J=2.3Hz, 1H), 7.44 (d, J=8.3 Hz, 1H), 7.25 (dd, J=2.2, 8.5 Hz, 1H), 6.95 (s,1H), 5.63 (d, J=3.5 Hz, 1H), 5.03 (s, 2H), 3.84 (d, J=3.5 Hz, 1H), 1.26(s, 9H); HPLC ret. time 2.37 min, 10-99% CH₃CN, 5 min run; ESI-MS 378.0m/z (MH⁺). Isomer B (yellow solid, 3.64 g, 23%, 96% de); ¹H NMR (400MHz, CDCl₃) □7.50 (d, J=2.1 Hz, 1H), 7.44 (d, J=8.3 Hz, 1H), 7.25 (dd,J=2.1, 8.5 Hz, 1H), 6.88 (d, J=0.5 Hz, 1H), 5.65 (d, J=2.5 Hz, 1H), 5.02(s, 2H), 3.88 (d, J=2.3 Hz, 1H), 1.26 (s, 9H); HPLC ret time 2.50 min,10-99% CH₃CN, 5 min run; ESI-MS 378.2 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(2-chlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamideand1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide

To 1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid (864 , mg 4.2mmol) was slowly added SOCl₂ (916 μL, 12.6 mmol) followed by DMF (3drops). The reaction mixture was heated at 60° C. for 0.5 h. The excessSOCl₃ was removed under vacuum. The acid chloride was then dissolved inanhydrous CH₂Cl₂ (6 mL) and was slowly added to a solution of(R)-N-((2-amino-4-methylthiazol-5-yl)(2-chlorophenyl)methyl)-2-methylpropane-2-sulfinamide(Isomer A) (1.35 g, 3.8 mmol) and Et₃N (2.92 mL, 21.0 mmol) in anhydrousCH₂Cl₂ (18 mL). The reaction mixture was stirred at room temperature for18 h, diluted with CH₂Cl₂ and washed with 1N HCl solution, saturatedaqueous NaHCO₃ solution and brine. The organic layer was dried overMgSO₄ and concentrated to provide1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) as an orange solid (1.54 g, 75%) that was used withoutfurther purification. HPLC ret time 3.59 min, 10-99% CH₃CN, 5 min run;ESI-MS 546.5 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer B), was made by the procedure used for Isomer A starting from1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid and(R)-N-(2-amino-4-methylthiazol-5-yl)(2-chlorophenyl)methyl)-2-methylpropane-2-sulfinamide(Isomer B). The crude product was adsorbed onto silica gel and purifiedby column chromatography (25-60% EtOAc/hexanes) to yield the product aslight orange solid (62%). ¹H NMR (400 MHz, CDCl₃) δ 8.48 (s, 1H), 7.70(dd, J=7.8, 1.6 Hz, 1H), 7.34-7.26 (m, 3H), 6.88-6.79 (m, 3H), 6.26 (d,J=1.9 Hz, 1H), 6.01 (s, 2H), 3.55 (d, J=1.7 Hz, 1H), 2.40 (s, 3H), 1.68(m, 2H), 1.28-1.18 (m, 11H; HPLC ret. time 3.69 min, 10-99% CH₃CN, 5 minrun; ESI-MS 546.5 m/z (MH⁺).

N-(5-((R)-(2-Chlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideandN-(5-((S)-(2-Chlorophenyl)((R)-1,1dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

N-(5-((2-Chlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer A), was made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) starting from1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxylic acid and(R)-N-((2-amino-4-methylthiazol-5-yl)(2-chlorophenyl)methyl)-2-methylpropane-2-sulfinamide(Isomer A). The crude product was adsorbed onto silica gel and purifiedby column chromatography (25-100% EtOAc/hexanes) to yield an orangesolid (55%). ¹H NMR (400 MHz, CDCl₃) δ 8.36 (s, 1H), 7.67 (dd, J=7.7,1.6 Hz, 1H), 7.35-7.21 (m, 3H), 7.17-7.07 (m, 3H), 6.21 (d, J=2.5 Hz,1H), 3.80 (d, J=2.4 Hz, 1H), 2.41 (s, 3H), 1.30-1.70 (m, 2H), 1.26-1.18(m, 11H); HPLC ret. time 3.81 min, 10-99% CH₃CN, 5 min run; ESI-MS 582.3m/z (MH⁺).

N-(5-((2-Chlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer B), was made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) starting from1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid and(R)-N-((2-amino-4-methylthiazol-5-yl)(2-chlorophenyl)methyl)-2-methylpropane-2-sulfinamide(Isomer B). The crude product was adsorbed onto silica gel and purifiedby column chromatography (30-80% EtOAc/hexanes) to yield an orange solid(37%) ¹H NMR (400 MHz, CDCl₃) δ 8.33 (s, 1H), 7.70 (dd, J=7.8, 1.7 Hz,1H), 7.35-7.20 (m, 3H), 7.16-7.07 (m, 3H), 6.26 (d, J=2.0 Hz, 1H), 3.55(d, J=1.8 Hz, 1H), 2.39 (s, 3H), 1.79-1.71 (m, 2H), 1.28-1.21 (m, 11H);HPLC ret. time 3.98 min, 10-99% CH₃CN, 5 min run; ESI-MS 582.3 m/z(MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(3,4-dichlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamideand1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(3,4-dichlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((3,4-dichlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide(Isomer A), was made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) starting from1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid, (COCl)₂ and(R)-N-((2-aminothiazol-5-yl)(3,4-dichlorophenyl)methyl)-2-methylpropane-2-sulfinamide(Isomer A). The crude product was purified by column chromatography(60-80% EtOAc/Hexane) to provide a yellow solid (2.70 g, 72%). ¹H NMR(400 MHz, CDCl₃) δ 8.57 (s, 1H), 7.49 (d, J=2.1 Hz, 1H), 7.43 (d, J=8.3Hz, 1H), 7.28-7.26 (m, 2H), 6.90 (dd, J=1.8, 7.9 Hz, 1H), 6.87 (d, J=1.6Hz, 1H), 6.82 (d, J=7.9 Hz, 1H), 6.02 (s, 2H), 5.74 (d, J=3.1 Hz, 1H),3.73 (d, J=3.2 Hz, 1H), 1.73-1.71 (m, 2H), 1.28-1.21 (m, 11H; HPLC rettime 3.80 min, 10-99% CH₃CN, 5 min run: ESI-MS 566.2 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((3,4-dichlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide(Isomer B), was made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) starting from1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid, (COCl)₂ and(R)-N-((2-aminothiazol-5-yl)(3,4-dichlorophenyl)methyl)-2-methylpropane-2-sulfinamide(Isomer B). The crude product was purified by column chromatography(60-100% EtOAc/Hexane) to provide a yellow solid (2.65 g, 71%). ¹H NMR(400 MHz, CDCl₃) δ 8.59 (s, 1H), 7.52 (d, J=2.1 Hz, 1H), 7.42 (d, J=8.3Hz, 1H), 7.29-7.26 (m, 2H), 6.90-6.86 (m, 2H), 6.81 (d, J=7.9 Hz, 1H),6.02 (dd, J=1.4, 1.9 Hz, 2H), 5.77 (d, J=1.8 Hz, 1H), 3.72 (d, J=1.9 Hz,1H), 1.72 (dd, J=3.2, 6.7 Hz, 2H), 1.28-1.22 (m, 11H); HPLC ret. time3.91 min, 10-99% CH₃CN, 5 min run; ESI-MS 566.4 m/z (MH⁺).

(R)-N-(5-(Amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideand(S)-N-(5-(Amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

To a solution of1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A), (1.47 g, 2.69 mmol) in MeOH (13 mL) was added 4 M HCl indioxane (4 mL, 16 mmol). The reaction mixture was stirred at roomtemperature for 3 h and then concentrated. The crude product wasdissolved in CH₂Cl₂ and washed with saturated aqueous NaHCO₃ solution(×2) and brine, then dried over MgSO₄ and concentrated. The crudeproduct was adsorbed onto silica get and purified by columnchromatography (70-100% EtOAc/hexanes) to provideN-(5-(amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide(Isomer A), as an orange solid (540 mg, 45%). ¹H NMR (400 MHz, DMSO-d₆)δ 7.80 (d, J=7.9 Hz, 1H), 7.40-7.36 (m, 2H), 7,27 (m, 1H, 6.94 (s, 1H),6.85 (m, 2H), 6.00 (s, 2H), 5.56 (s, 1H), 2.19 (s, 3H), 1.40 (m, 2H),1.10 (m, 2H); HPLC ret. time 2.65 min, 10-99% CH₃CN 5 min run; ESI-MS442.5 m/z (MH⁺).

N-(5-(Amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer B) was made by the procedure used for Isomer A starting from1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer B) (90%). ¹H NMR (400 MHz, DMSO-d6) δ 7.80 (d, J=7.8 Hz, 1H),7.40-7.36 (m, 2H), 7.27 (m, 1H), 6.94 (s, 1H), 6.85 (m, 2H), 6.00 (s,2H), 5.56 (s, 1H), 2.19 (s, 3H), 1.40 (m, 2H), 1.10 (m, 2H); HPLC ret.time 2.69 min, 10-99% Ch₃CN, 5 min run; ESI-MS 442.3 m/z (MH⁺).

(R)-N-(5-(Amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(2,2-(difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideand(S)-N-(5-(Amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(2,2-(difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

N-(5-(Amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(2,2-(difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer A) was made by the procedure used forN-(5-(amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer A) starting fromN-(5-((2-chlorophenyl)((R)-1,1dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer A) (83%). ¹H NMR (400 MHz, DMSO-d6) δ 7.81 (m, 1H), 7.41-7.32(m, 4H), 7.27 (m, 1H), 7.18 (dd, J=1.7, 8.3 Hz, 1H), 5.57 (s, 1H), 2.20(s, 3H), 1.48 (m, 2H), 1.17 (m, 2H); HPLC ret. time 2.84 min, 10-99%Ch₃CN, 5 min run; ESI-MS 478.1 m/z (MH⁺).

N-(5-(Amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer B) was made by the procedure used forN-(5-(amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer A) starting fromN-(5-(2-chlorophenyl)((R)-1,1dimethylethylsulfinamido)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer B) (65%). ¹H NMR (400 MHz, DMSO-d6) δ 7.81 (m, 1H), 7.41-7.32(m, 4H), 7.28 (m, 1H), 7.18 (dd, J=1.7, 8.3 Hz, 1H), 5.56 (s, 1H), 2.19(s, 3H), 1.48 (m, 2H), 1.18 (m, 2H); HPLC ret. time 2.84 min 10-99%CH₃CN, 5 min run; ESI-MS 478.1 m/z (MH⁺).

(R)-N-(5-(Amino(3,4-dichlorophenyl)methyl)-thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideand(S)-N-(5-(Amino(3,4-dichlorophenyl)methyl)-thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

N-(5-(Amino(3,4-dichlorophenyl)methyl)-thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer A) was made by the procedure used forN-(5-(amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer A) starling from1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((3,4-dichlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide(Isomer A). The crude product was purified by column chromatography(0-20% MeOH/DCM) (98%). ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (bs, 1H), 7.70(d, J=2.0 Hz, 1H), 7.58 (d, J=8.3 Hz, 1H), 7.40 (dd, J=2.0, 8.4 Hz, 1H),7.16 (d, J=0.8 Hz, 1H), 6.96 (s, 1Hi, 6.86 (d, J=0.9 Hz, 2H), 6.01 (s,2H), 5.29 (s, 1H), 1.44-1.42 (m, 2H), 1.13-1.10 (m, 2H); HPLC ret. time2.81 min, 10-99% CH₃CN, 5 min run; ESI-MS 462.3 m/z (MH⁺).

N-(5-(Amino(3,4-dichlorophenyl)methyl)-thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer B) was made by the procedure used for (Isomer A) starting from1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((3,4-dichlorophenyl)((R)-1,1-dimethylethylsulfinamido)methyl)thiazol-2-yl)cyclopropanecarboxamide(Isomer B) (quant.). ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (bs, 1H), 7.70 (d,J=2.0 Hz, 1H), 7.58 (d, J=8.3 Hz, 1H), 7.40 (dd, J=2.0, 8.4 Hz, 1H),7.16 (d, J=0.7 Hz, 1H), 6.96 (s, 1H), 6.86 (d, J=0.9 Hz, 2H), 6.01 (s,2H), 5.29 (s, 1H), 1.44-1.42 (m, 2H), 1.13-1.10 (m, 2H); HPLC ret. time2.81 min, 10-99% CH₃CN, 5 min run; ESI-MS 462.1 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-((S)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamideand1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-((S)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide

To a solution ofN-(5-(amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer A) (450 mg, 1.02 mmol) in MeOH (5 mL) was added(S)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanal (289 mg, 1.22 mmol).The reaction mixture was stirred at room temperature for 20 min beforeNaBH₄ (58 mg, 1.53 mmol) was added. Stirring was continued at roomtemperature for 3 h. After approximately 1 h some precipitate/gumstarted to form in the reaction solution so MeOH (5 mL) and CH₂Cl₂ (2mL) were added to keep everything in solution. The reaction was dilutedwith H₂O and extracted with EtOAc. The combined organic layers waswashed with brine, dried over MgSO₄ and concentrated. The residue wasadsorbed onto silica gel and purified by column chromatography (0-25%EtOAc/hexanes) to afford1-(benzo[d][1,3]dioxol-5-yl)-N-(5-(((S)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) as a pale yellow solid (470 mg, 74%). ¹H NMR (400 MHz, CDCl₃)δ 8.40 (s, 1H), 7.83 (dd, J=8.1, 1.6 Hz, 1H), 7.28-7.25 (m, 2H), 7.13(m, 1H), 6.88-6.83 (m, 2H), 6.79 (d, J=7.9 Hz, 1H), 6.00 (s, 2H), 5.08(s, 1H), 4.33 (m, 1H), 2.94 (dd, J=9.9, 6.3 Hz, 1H), 2.67 (td, J=8.3,3.4 Hz, 1H), 2.53 (q, J=8.4 Hz, 1H), 2.37 (s, 3H), 2.17 (dd, J=9.9, 4.7Hz, 1H), 2.00 (m, 1H), 1.74-1.61 (m, 3H), 1.17 (m, 2H), 0.86 (m, 9H),0.01 (s, 3H), −0.01 (s, 3H); HPLC ret. time 3.45 min, 10-99% CH₃CN, 5min run; ESI-MS 626.5 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer B) was made by the procedure used, for (Isomer A) starting fromN-(5-(amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer B) and (S)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanal (81%).¹H NMR (400 MHz, CDCl₃) δ 8.41 (s, 1H), 7.86 (dd, J=7.2, 0.7 Hz, 1H),7.27-7.24 (m, 2H), 7.12 (m, 1H), 6.88-6.84 (m, 2H), 6.79 (d, J=7.9 Hz,1H), 6.00 (s, 2H), 5.05 (s, 1H), 4.34 (m, 1H), 2.84 (dd, J=9.7, 6.3 Hz,1H), 2.56-2.46 (m, 2H), 2.37-2.32 (m, 4H), 2.03 (m, 1H), 1.74-1.62 (m,3H), 1.18 (m, 2H), 0.86 (m, 9H), 0.02 (s, 3H), 0.00 (s, 3H); HPLC ret.time 3.54 min, 10-99% CH₃CN, 5 min run; ESI-MS 626.3 m/z (MH⁺).

N-(5-((R)-((S)-3-(tert-Butyldimethylsilyloxy)pyrrolidin-1yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamideandN-(5-((S)-((S)-3-(tert-Butyldimethylsilyloxy)pyrrolidin-1yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide

N-(5-(((S)-3-(tert-Butyldimethylsilyloxy)pyrrolidin-1yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide(Isomer A) was made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5(((S)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) starting fromN-(5-(amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer A) and (S)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanal (73%).¹H NMR (400 MHz, CDCl₃) δ 8.25 (s, 1H), 7.83 (dd, J=8.1, 1.6 Hz, 1H),7.28-7.25 (m, 2H), 7.16-7.11 (m, 3H), 7.06 (d, J=8.1 Hz, 1H), 5.08 (s,1H), 4.33 (m, 1H), 2.94 (dd, J=9.9, 6.2 Hz, 1H), 2.66 (td, J=8.3, 3.7Hz, 1H), 2.54 (q, J=8.3 Hz, 1H), 2.36 (s, 3H), 2.18 (dd, J=9.9, 4.7 Hz,1H), 2.02 (m, 1H), 1.75 (m, 2H), 1.65 (m, 1H), 1.20 (m, 3H), 0.86 (m,9H), 0.01 (s, 3H), −0.01 (s, 3H); HPLC ret. time 3.69 min, 10-99% CH₃CN,5 min run; ESI-MS 662.1 m/z (MH⁺).

N-(5-(((S)-3-(tert-Butyldimethylsilyloxy)pyrrolidin-1yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide(Isomer B) was made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-(((S)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) starting fromN-(5-(amino(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer B) and (S)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanal (93%).¹H NMR (400MHz, CDC₃) δ 8.26 (ss 1H, 7.85 (dd, J=8.1, 1.7 Hz, 1H),7.28-7.24 (m, 2H), 7.16-7.10 (m, 3H), 7.07 (d, J=8.2 Hz, 1H), 5.05 (s,1H), 4.34 (m, 1H), 2.83 (dd, J=9.7, 6.3 Hz, 1H), 2.55-2.47 (m, 2H),2.36-2.33 (m, 4H), 2.04 (m, 1H), 1.79-1.63 (m, 3H), 1.21 (m, 2H), 0.86(m, 9H), 0.02 (s, 3H), 0.00 (s, 3H); HPLC ret. time 3.73 min, 10-99%CH₃CN, 5 min run; ESI-MS 662.1 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(3,4-dichlorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamideand1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-((R)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(3,4-dichlorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-(((R)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(3,4-dichlorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide(Isomer A) was made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-(((S)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) starting fromN-(5-(amino(3,4-dichlorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer A) and (R)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanal (85%).¹H NMR (400 MHz, CDCl₃) δ 8.47 (s, 1H), 7.51 (d, J=2.0 Hz, 1H), 7.35 (d,J=8.3 Hz, 1H), 7.30 (dd, J=2.0, 8.3 Hz, 1H), 7.19 (s, 1H), 6.90-6.85 (m,2H), 6.80 (d, J=7.9 Hz, 1H), 6.01 (s, 2H), 4.47 (s, 1H), 4.34 (m, 1H),2.84 (dd, J=6.2, 9.8 Hz, 1H), 2.55-2.44 (m, 2H), 2.31 (dd, J=4.4, 9.9Hz, 1H), 2.17 (s, 1H), 1.73-1.64 (m, 3H), 1.26-1.19 (m, 2H), 0.91 (s,3H), 0.86 (s, 6H), 0.02 (s, 6H); HPLC ret. time 3.66 min, 10-99% CH₃CN,5 min run; ESI-MS 646.5 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-(((R)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(3,4-dichlorophenyl)methyl)thiazol-2-yl)cyclopropanecarboxamide(Isomer B) was made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-(((S)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) starting fromN-(5-(amino(3,4-dichlorophenyl)methyl)thiazol-2-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer B) and (R)-3-(tert-butyldimethylsilyloxy)-4-chlorobutanal (70%).¹H NMR (400 MHz, CDCl₃) δ 8.47 (s, 1H), 7.50 (d, J=2.0 Hz, 1H), 7.36 (d,J=8.3 Hz, 1H), 7.29 (dd, J=2.0, 8.3 Hz, 1H), 7.19 (s, 1H), 6.90-6.85 (m,2H), 6.80 (d, J=7.9 Hz, 1H), 6.01 (s, 2H), 4.51 (s, 1H), 4.34 (m, J=3.4,9.4 Hz, 1H), 2.88 (dd, J=6.2, 10.0 Hz, 1H), 2.6-2.62 (m, 1H), 2.53 (dd,J=8.2, 16.7 Hz, 1H), 2.17 (dd, J=4.4, 10.0 Hz, 1H), 2.07-1.78 (m, 1H),1.75-1.63 (m, 3H), 1.22-1.18 (m, 2H), 0.86 (d, J=2.5 Hz, 9H), 0.01 (m,6H); HPLC ret. time 3.67 min, 10-99% CH₃CN, 5 min run; ESI-MS 646.4 m/z(MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(2-chlorophenyl)((S)-3-hydroxypyrrolidin-1-yl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamideand1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(2-chlorophenyl)((S)-3-hydroxypyrrolidin-1-yl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide

A mixture of1-(benzo[d][1,3]dioxol-5-yl)-N-(5-(((S)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) (400 mg, 0.64 mmol) and TBAF (1M in THF, 3.84 mL, 3.84 mmol)was stirred at room temperature for 5 h. The reaction was partitionedbetween H₂O and EtOAc. The combined organic layers were washed withbrine, dried over MgSO₄ and concentrated. The residue was adsorbed ontosilica gel and purified by column chromatography (25-75% EtOAc/hexanes)to afford1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((S)-3-hydroxypyrrolidin-1-yl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) as a pale yellow solid (245 mg, 75%, >99% de). ¹H NMR (400MHz, CDCl₃) δ 8.41 (s, 1H), 7.85 (dd, J=8.1, 1.7 Hz, 1H), 7.29-7.26 (m,2H), 7.14 (m, 1H), 6.88-6.84 (m, 2H), 6.79 (d, J=7.9 Hz, 1H), 6.00 (s,2H), 5.07 (s, 1H), 4.30 (m, 1H), 2.79 (td, J=8.6, 5.8 Hz, 1H), 2.63-2.56(m, 2H), 2.36 (s, 3H), 2.29 (td, J=9.0, 5.9 Hz, 1H), 2.15 (m, 1H), 1.85(d, J=7.9 Hz, 1H), 1.78-1.65 (m, 3H), 1.18 (m, 2H); HPLC ret. time 2.70min, 10-99% CH₃CN, 5 min run; ESI-MS 512.5 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((S)-3-hydroxypyrrolidin-1-yl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer B) was made by the procedure used for (Isomer A) starting from1-(benzo[d][1,3]dioxol-5-yl)-N-(5-(((S)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer B) (50%, >99% de). ¹H NMR (400 MHz, CDCl₃) δ 8.44 (s, 1H), 7.84(dd, J=7.8, 1.5 Hz, 1H), 7.29-7.25 (m, 2H), 7.14 (td, J=7.6, 1.7 Hz,1H), 6.88-6.84 (m, 2H), 6.79 (d, J=7.9 Hz, 1H), 6.00 (s, 2H), 5.07 (s,1H), 4.29 (m, 1H), 2.83 (td, J=8.6, 5.0 Hz, 1H), 2.61 (d, J=10.1 Hz,1H), 2.49 (m, 1H), 2.36-2.29 (m, 4H), 2.17 (m, 1H), 1.89 (d, J=7.9 Hz,1H), 1.76-1.66 (m, 3H), 1.19 (m, 2H); HPLC ret. time 2.71 min, 10-99%CH₃CN, 5 min run; ESI-MS 512.5 m/z (MH⁺).

N-(5-((R)-(2-Chlorophenyl)((S)-3-hydroxypyrrolidin-1-yl)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamideandN-(5-((S)-(2-Chlorophenyl)((S)-3-hydroxypyrrolidin-1-yl)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide

N-(5-((2-Chlorophenyl)((S)-3-hydroxypyrrolidin-1-yl)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer A) was made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((S)-3-hydroxypyrrolidin-1-yl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) starting fromN-(5-(((S)-3-(tert-Butyldimethylsilyloxy)pyrrolidin-1yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide(Isomer A) (72%). ¹H NMR (400 MHz, CDCl₃) δ 8.28 (s, 1H), 7.84 (dd,J=8.1, 1.7 Hz, 1H), 7.30-7.26 (m, 2H), 7.17-7.12 (m, 3H), 7.07 (d, J=8.2Hz, 1H), 5.07 (s, 1H), 4.30 (m, 1H), 2.79 (td, J=8.6, 5.7 Hz, 1H),2.62-2.55 (m, 2H), 2.35 (s, 3H), 2.28 (td, J=9.0, 5.9 Hz, 1H), 2.15 (m,1H), 1.86 (d, J=7.9 Hz, 1H), 1.80-1.71 (m, 3H), 1.21 (m, 2H); HPLC ret.time 2.91 min, 10-99% CH₃CN, 5 min run; ESI-MS 548.3 m/z (MH⁺).

N-(5-((2-Chlorophenyl)((S)-3-hydroxypyrrolidin-1-yl)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide(Isomer B) was made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((2-chlorophenyl)((S)-3-hydroxypyrrolidin-1-yl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) starting fromN-(5-(((S)-3-(tert-Butyldimethylsilyloxy)pyrrolidin-1yl)(2-chlorophenyl)methyl)-4-methylthiazol-2-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide(Isomer B) (85%). ¹H NMR (400 MHz, CDCl₃) δ 8.28 (s, 1H), 7.82 (dd,J=7.7, 1.5 Hz, 1H), 7.30-7.25 (m, 2H), 7.14 (m, 3H), 7.07 (d, J=8.2 Hz,1H), 5.07 (s, 1H), 4.29 (m, 1H), 2.83 (td, J=8.7, 5.1 Hz, 1H), 2.61 (d,J=10.0 Hz, 1H), 2.50 (dd, J=10.1, 5.1 Hz, 1H), 2.36-2.29 (m, 4H), 2.17(m, 1H), 1.88 (d, J=7.8 Hz, 1H), 1.82-1.70 (m, 3H), 1.22 (m, 2H); HPLCret. time 2.94 min, 10-99% CH₃CN, 5 min run; ESI-MS 548.3 m/z (MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(3,4-dichlorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamideand1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((S)-(3,4-dichlorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(3,4-dichlorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide(Isomer A) was made by the procedure used for1-(benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(3,4-dichlorophenyl)((S)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide(Isomer A) starting from1-(benzo[d][1,3]dioxol-5-yl)-N-(5-(((R)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(3,4-dichlorophenyl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer A) The crude product was purified by column chromatography(60-90% EtOAc/hexane) (74%, 98% de). ¹H NMR (400 MHz, CDCl₃) δ 8.49 (s,1H), 7.50 (d, J=2.0 Hz, 1H), 7.37 (d, J=8.3 Hz, 1H), 7.31 (dd, J=2.0,8.3 Hz, 1H), 7.20 (s, 1H), 6.90-6.86 (m, 2H), 6.81 (d, J=7.9 Hz, 1H),6.01 (s, 2H), 4.48 (s, 1H), 4.32 (m, 1H), 2.82-2.76 (m, 1H), 2.59-2.57(m, 1H), 2.51 (q, J=5.1 Hz, 1H), 2.33 (td, J=8.9, 6.1 Hz, 1H), 2.16 (dd,J=5.3, 12.5 Hz, 1H), 1.81-1.69 (m, 4H), 1.22 (dd, J=5.0, 7.6 Hz, 2H);HPLC ret. time 2.81 min, 10-99% CH₃CN, 5 min run; ESI-MS 532.2 m/z(MH⁺).

1-(Benzo[d][1,3]dioxol-5-yl)-N-(5-((R)-(3,4-dichlorophenyl)((R)-3-hydroxypyrrolin-1-yl)methyl)thiazol-2-yl)cyclopropanecarboxamide(Isomer B) was made by the procedure used for (Isomer A) starting from1-(benzo[d][1,3]dioxol-5-yl)-N-(5-(((R)-3-(tert-butyldimethylsilyloxy)pyrrolin-1-yl)(3,4-dichlorophenyl)methyl)-4-methylthiazol-2-yl)cyclopropanecarboxamide(Isomer B) (78%, 95% de). ¹H NMR (400 MHz, CDCl₃) δ 8.49 (s, 1H), 7.52(d, J=2.0 Hz, 1H), 7.37 (d, J=8.3 Hz, 1H), 7.30 (dd, J=2.0, 8.3 Hz, 1H),7.20 (s, 1H), 6.90-6.86 (m, 2H), 6.81 (d, J=7.9 Hz, 1H), 6.01 (s, 2H),4.48 (s, 1H), 4.34-4.32 (m, 1H), 2.76 (td, J=8.5, 6.0 Hz, 1H), 2.57 (m,2H), 2.29 (m, 1H), 2.15 (q, J=7.2 Hz, 1H), 1.81-1.68 (m, 4H), 1.25-1.20(m, 2H); HPLC ret. time 2.80 min, 10-99% CH₃CN, 5 min run; ESI-MS 532.2m/z (MH⁺).

Assays For Detecting and Measuring ΔF508-CFTR Correction Properties ofCompounds

I) Membrane Potential Optical Methods For Assaying ΔF508-CFTR ModulationProperties of Compounds

The optical membrane potential assay utilized voltage-sensitive FRETsensors described by Gonzalez and Tsien (See, Gonzalez, J. E, and R. Y.Tsien (1995) “Voltage sensing by fluorescence resonance energy transferin single cells” Biophys J 69(4): 1272-80, and Gonzalez, J. E. and R. Y.Tsien (1997) “Improved indicators of cell membrane potential that usefluorescence resonance energy transfer” Chem. Biol 4(4): 269-77) incombination with instrumentation for measuring fluorescence changes suchas the Voltage/Ion Probe Reader (VIPR) (See, Gonzalez, J. E., K. Oades,et al. (1999) “Cell-based assays and instrumentation for screeningion-channel targets” Drug Discov Today 4(9): 431-439).

These voltage sensitive assays are based on the change in fluorescenceresonant energy transfer (FRET) between the membrane-soluble,voltage-sensitive dye, DiSBAC₂(3), and a fluorescent phospholipid,CC2-DMPE, which is attached to the outer leaflet of the plasma membraneand acts as a FRET donor. Changes in membrane potential (V_(m)) causethe negatively charged DiSBAC₂(3) to redistribute across the plasmamembrane and the amount of energy transfer from CC2-DMPE changesaccordingly. The changes in fluorescence emission were monitored usingVIPR™ II, which is an integrated liquid handler and fluorescent detectordesigned to conduct cell-based screens in 96- or 384-well microtiterplates.

Cell Culture

NIH3T3 mouse fibroblasts stably expressing ΔF508-CFTR are used foroptical measurements of membrane potential. The cells are maintained at37° C. in 5% CO₂ and 90% humidity in Dulbecco's modified Eagle's mediumsupplemented with 2 mM glutamine, 10% fetal bovine serum, 1× NEAA, β-ME,1× pen/strep, and 25 mM HEPES in 175 cm² culture flasks. For all opticalassays, the cells were seeded at 30,000/well in 384-well matrigel-coatedplates and cultured for 2 hrs at 37° C. before culturing at 27° C. for24 hrs. for the potentiator assay. For the correction assays, the cellsare cultured at 27° C. or 37° C. with and without compounds for 16-24hoursB) Electrophysiological Assays for assaying ΔF508-CFTR modulationproperties of compounds

1. Ussing Chamber Assay

Ussing chamber experiments were performed on polarized epithelial cellsexpressing ΔF508-CFTR to further characterize the ΔF508-CFTR modulatorsidentified in the optical assays. FRT^(ΔF508-CFTR) epithelial cellsgrown on Costar Snapwell cell culture inserts were mounted in an Ussingchamber (Physiologic Instruments, Inc., San Diego, Calif.), and themonolayers were continuously short-circuited using a Voltage-clampSystem (Department of Bioengineering, University of Iowa, IA, and,Physiologic Instruments, inc., San Diego, Calif.). Transepithelialresistance was measured by applying a 2-mV pulse. Under theseconditions, the FRT epithelia demonstrated resistances of 4 KΩ/cm² ormore. The solutions were maintained at 27° C. and bubbled with air. Theelectrode offset potential and fluid resistance were corrected using, acell-free insert. Under these conditions, the current reflects the flowof Cl⁻ through ΔF508-CFTR expressed in the apical membrane. The I_(SC)was digitally acquired using an MP100A-CE interface and AcqKnowledgesoftware (v3.2.6; BIOPAC Systems, Santa Barbara, Calif.).

Identification of Correction Compounds

Typical protocol utilized a basolateral to apical membrane Cl⁻concentration gradient. To set up this gradient, normal ringer was usedon the basolateral membrane, whereas apical NaCl was replaced byequimolar sodium gluconate (titrated to pH 7.4 with NaOH) to give alarge Cl⁻ concentration gradient across the epithelium. All experimentswere performed with intact monolayers. To fully activate ΔF508-CFTR,forskolin (10 μM) and the PDE inhibitor, IBMX (100 μM), were appliedfollowed by the addition of the CFTR potentiator, genistein (50 μM).

As observed in other cell types, incubation at low temperatures of FRTcells stably expressing ΔF508-CFTR increases the functional density ofCFTR in the plasma membrane. To determine the activity of correctioncompounds, the cells were incubated with 10 μM of the test compound for24 hours at 37° C. and were subsequently washed 3× prior to recording.The cAMP- and genistein-mediated I_(SC) in compound-treated cells wasnormalized to the 27° C. and 37° C. controls and expressed as percentageactivity. Preincubation of the cells with the correction compoundsignificantly increased the cAMP- and genistem-mediated I_(SC) comparedto the 37° C. controls.

To determine the activity of correction compounds for increasing thedensity of functional ΔF508-CFTR in the plasma membrane, we used theabove-described perforated-patch-recording techniques to measure thecurrent density following 24-hr treatment with the correction compounds.To fully activate ΔF508-CFTR, 10 μM forskolin and 20 μM genistein wereadded to the cells. Under our recording conditions, the current densityfollowing 24-hr incubation at 27° C. was higher than that observedfollowing 24-hr incubation at 37° C. These results are consistent withthe known effects of low-temperature incubation on the density ofΔF508-CFTR in the plasma membrane. To determine the effects ofcorrection compounds on CFTR current density, the cells were incubatedwith 10 μM of the test compound for 24 hours at 37° C. and the currentdensity was compared to the 27° C. and 37° C. controls (% activity).Prior to recording, the cells were washed 3× with extracellularrecording medium to remove any remaining test compound. Preincubationwith 10 μM of correction compounds significantly increased the cAMP- andgenistein-dependent current compared to the 37° C. controls.

Table 3 illustrates the EC50 and relative efficacy of exemplaryembodiments of the present invention. In Table 3, the following meaningsapply:

-   EC50: “+++” means <2 uM; “++” means between 2 uM to 20 uM; “+” means    between 25 uM to 60 uM.-   % Efficacy: “+” means <25%; “++” means between 25% to 100%; “+++”    means >100%.

TABLE 3 Cmpd No. EC 50 % Efficacy 1 +++ +++ 2 ++ +++ 3 +++ +++ 4 +++ +++5 +++ +++ 6 +++ +++ 7 +++ +++ 8 +++ +++ 9 +++ +++ 10 +++ +++ 11 +++ +++12 +++ ++ 13 +++ +++As illustrated in Table 3 above, the compounds of the present inventionexhibit unexpectedly better correction activity as measured by theassays above.

1-21. (canceled)
 22. A method of increasing the number of functionalCFTR in a membrane of a cell, comprising the step of contacting saidcell with a compound according to a compound of formula I or formula II:

or a pharmaceutically acceptable salt thereof, wherein: each R^(X) isindependently hydrogen, halo, CF₃, C₁-C₄ alkyl, or —OC₁-C₄ alkyl;provided that both R^(X) are not simultaneously hydrogen: or the twoR^(X), taken together form ring (a):

X is CH₂, CF₂, CH₂—CH₂, or CF₂—CF₂; ring A is 3-7 membered monocycliccycloalkyl ring; R^(AA) and R^(BB) taken together with the nitrogenatom, form a pyrrolidinyl ring substituted with OR′; R′ is hydrogen orC₁-C₆ aliphatic, wherein up to two carbon units of said aliphatic areoptionally and independently replaced by —CO—, —CS—, —COCO—, —CONR—,—CONRNR—, —CO₂—,—OCO—, —NRCO₂—, —O—, —NRCONR—, —OCONR—, —NRNR, —NRNRCO—,—NRCO—, —S—, —SO, —SO₂—, —NR—, —SO₂NR—, NRSO₂—, or —NRSO₂NR—; R ishydrogen or C₁-C₆ aliphatic; Z is halo, CN, COOH, CF₃, ordifluoromethylenedioxy; and q is 0-3. 23-66. (canceled)
 67. The methodof claim 22, wherein two R^(X) taken together form ring (a) and X isCH₂.
 68. The method of claim 22, wherein wherein two R^(X) takentogether form ring (a) and X is CF₂.
 69. The method of claim 22, whereinone R^(X) is hydrogen and the other R^(X) is halo, CF₃, C₁-C₄ alkyl, or—OC₁-C₄ alkyl.
 70. The method of claim 22, wherein one R^(X) is hydrogenand the other R^(X) is 4-methoxy.
 71. The method of claim 22, whereinring A is cyclopropyl, cyclopentyl, or cyclohexyl.
 72. The method ofclaim 71, wherein ring A is cyclopropyl or cyclopentyl.
 73. The methodof claim 71, wherein ring A is cyclopropyl.
 74. The method of claim 22,wherein R′ is hydrogen.
 75. The method of claim 22, wherein R′ is C₁₋₆alkyl.
 76. The method of claim 22, wherein R^(AA) and R^(BB), takentogether, form a pyrrolidinyl with an OH substituent.
 77. The method ofclaim 22, wherein Z is selected from halo, CF₃, ordifluoromethylenedioxy.
 78. The method of claim 22, wherein q is
 0. 79.The method of claim 22, wherein q is
 1. 80. The method of claim 22,wherein R is hydrogen.
 81. The method of claim 22, wherein compounds offormula I or formula II comprise one or more of the following features:two R^(X) taken together form ring (a); X is CH₂; ring A is cyclopropyl;R is hydrogen; q is 1; and Z is halo, CF₃, or difluoromethylenedioxy.82. The method of claim 22, wherein R^(AA) and R^(BB) in formula I,taken together with the nitrogen atom, form the following ring (i):


83. The method of claim 22, wherein R^(AA) and R^(BB) in formula I,taken together with the nitrogen atom, form the following ring (ii):


84. The method of claim 22, wherein the compound is selected from